Isolation and characterization of two cDNA clones encoding for glutamate dehydrogenase in Nicotiana plumbaginifolia.

Science.gov (United States)

Ficarelli, A; Tassi, F; Restivo, F M

1999-03-01

We have isolated two full length cDNA clones encoding Nicotiana plumbaginifolia NADH-glutamate dehydrogenase. Both clones share amino acid boxes of homology corresponding to conserved GDH catalytic domains and putative mitochondrial targeting sequence. One clone shows a putative EF-hand loop. The level of the two transcripts is affected differently by carbon source.

A new approach for cloning hLIF cDNA from genomic DNA isolated from the oral mucous membrane.

Science.gov (United States)

Cui, Y H; Zhu, G Q; Chen, Q J; Wang, Y F; Yang, M M; Song, Y X; Wang, J G; Cao, B Y

2011-11-25

Complementary DNA (cDNA) is valuable for investigating protein structure and function in the study of life science, but it is difficult to obtain by traditional reverse transcription. We employed a novel strategy to clone human leukemia inhibitory factor (hLIF) gene cDNA from genomic DNA, which was directly isolated from the mucous membrane of mouth. The hLIF sequence, which is 609 bp long and is composed of three exons, can be acquired within a few hours by amplifying each exon and splicing all of them using overlap-PCR. This new approach developed is simple, time- and cost-effective, without RNA preparation or cDNA synthesis, and is not limited to the specific tissues for a particular gene and the expression level of the gene.

Isolation and characterization of human cDNA clones encoding the α and the α' subunits of casein kinase II

International Nuclear Information System (INIS)

Lozeman, F.J.; Litchfield, D.W.; Piening, C.; Takio, Koji; Walsh, K.A.; Krebs, E.G.

1990-01-01

Casein kinase II is a widely distributed protein serine/threonine kinase. The holoenzyme appears to be a tetramer, containing two α or α' subunits (or one of each) and two β subunits. Complementary DNA clones encoding the subunits of casein kinase II were isolated from a human T-cell λgt 10 library using cDNA clones isolated from Drosophila melanogasten. One of the human cDNA clones (hT4.1) was 2.2 kb long, including a coding region of 1176 bp preceded by 156 bp (5' untranslated region) and followed by 871 bp (3' untranslated region). The hT4.1 close was nearly identical in size and sequence with a cDNA clone from HepG2 human hepatoma cultured cells. Another of the human T-cell cDNA clones (hT9.1) was 1.8 kb long, containing a coding region of 1053 bp preceded by 171 by (5' untranslated region) and followed by 550 bp (3' untranslated region). Amino acid sequences deduced from these two cDNA clones were about 85% identical. Most of the difference between the two encoded polypeptides was in the carboxy-terminal region, but heterogeneity was distributed throughout the molecules. Partial amino acid sequence was determined in a mixture of α and α' subunits from bovine lung casein kinase II. The bovine sequences aligned with the 2 human cDNA-encoded polypeptides with only 2 discrepancies out of 535 amino acid positions. This confirmed that the two human T-cell cDNA clones encoded the α and α' subunits of casein kinase II. These studies show that there are two distinct catalytic subunits for casein II (α and α') and that the sequence of these subunits is largely conserved between the bovine and the human

Sequence of human protamine 2 cDNA

Energy Technology Data Exchange (ETDEWEB)

Domenjoud, L; Fronia, C; Uhde, F; Engel, W [Universitaet Goettingen (West Germany)

1988-08-11

The authors report the cloning and sequencing of a cDNA clone for human protamine 2 (hp2), isolated from a human testis cDNA library cloned in the vector {lambda}-gt11. A 66mer oligonucleotide, that corresponds to an amino acid sequence which is highly conserved between hp2 and mouse protamine 2 (mp2) served as hybridization probe. The homology between the amino acid sequence deduced from our cDNA and the published amino acid sequence for hp2 is 100%.

Transcriptomic survey of the midgut of Anthonomus grandis (Coleoptera: Curculionidae).

Science.gov (United States)

Salvador, Ricardo; Príncipi, Darío; Berretta, Marcelo; Fernández, Paula; Paniego, Norma; Sciocco-Cap, Alicia; Hopp, Esteban

2014-01-01

Anthonomus grandis Boheman is a key pest in cotton crops in the New World. Its larval stage develops within the flower bud using it as food and as protection against its predators. This behavior limits the effectiveness of its control using conventional insecticide applications and biocontrol techniques. In spite of its importance, little is known about its genome sequence and, more important, its specific expression in key organs like the midgut. Total mRNA isolated from larval midguts was used for pyrosequencing. Sequence reads were assembled and annotated to generate a unigene data set. In total, 400,000 reads from A. grandis midgut with an average length of 237 bp were assembled and combined into 20,915 contigs. The assembled reads fell into 6,621 genes models. BlastX search using the NCBI-NR database showed that 3,006 unigenes had significant matches to known sequences. Gene Ontology (GO) mapping analysis evidenced that A. grandis is able to transcripts coding for proteins involved in catalytic processing of macromolecules that allows its adaptation to very different feeding source scenarios. Furthermore, transcripts encoding for proteins involved in detoxification mechanisms such as p450 genes, glutathione-S-transferase, and carboxylesterases are also expressed. This is the first report of a transcriptomic study in A. grandis and the largest set of sequence data reported for this species. These data are valuable resources to expand the knowledge of this insect group and could be used in the design of new control strategies based in molecular information. © The Author 2014. Published by Oxford University Press on behalf of the Entomological Society of America.

Construction and characterization of cDNA library for IRM-2 mice

International Nuclear Information System (INIS)

Wang Qin; Li Jin; Song Li; Liu Qiang; Yue Jingyin; Mu Chuanjie; Tang Weisheng; Fan Feiyue

2010-01-01

Objective: To screen and isolate the radioresistance related genes of IRM-2 mice. Methods: cDNA library of IRM-2 mice was constructed by SMART technique. Total RNA was isolated from spleens of IRM-2 male mice. The first-strand cDNA was synthesized by using PowerScript reverse transcriptase, and double-strand cDNA was synthesized and amplified by long PCR. The PCR products were purified, digested with restriction enzyme Sfi I. The ds-cDNA fragment less than 500 bp was fractionated and ligated to the Sfi I-digested pDNR-LIB vector. The ligation mixture was transformed into E. coil DH5 α by electroporation transformation to generate the unamplified cDNA library. The quality of cDNA library was identified by PCR technique. 130 clones from cDNA library were sequenced and compared with GenBank database. Results: The cDNA library contained 2.25 x 10 6 independent clones with an average insert size of 1.2 kb. The ratio of recombination and full-length was 95% and 55%, respectively. 21 pieces of EST sequences from cDNA library were not the same as the known mice genes and registered into GenBank EST database, with registered number DW474856-DW474876. Conclusions: cDNA library of IRM-2 mice has been constructed successfully. 21 pieces of EST implies that radioresistance correlative genes may be in IRM-2 mice, which will lay a foundation for isolating and identifying radioresistance related genes in further study. (authors)

Isolation of CYP3A5P cDNA from human liver: a reflection of a novel cytochrome P-450 pseudogene.

Science.gov (United States)

Schuetz, J D; Guzelian, P S

1995-03-14

We have isolated, from a human liver cDNA library, a 1627 bp CYP3A5 cDNA variant (CYP3A5P) that contains several large insertions, deletions, and in-frame termination codons. By comparison with the genomic structure of other CYP3A genes, the major insertions in CYP3A5P cDNA demarcate the inferred sites of several CYP3A5 exons. The segments inserted in CYP3A5P have no homology with splice donor acceptor sites. It is unlikely that CYP3A5P cDNA represents an artifact of the cloning procedures since Southern blot analysis of human genomic DNA disclosed that CYP3A5P cDNA hybridized with a DNA fragment distinct from fragments that hybridized with either CYP3A5, CYP3A3 or CYP3A4. Moreover, analysis of adult human liver RNA on Northern blots hybridized with a CYP3A5P cDNA fragment revealed the presence of an mRNA with the predicted size of CYP3A5P. We conclude that CYP3A5P cDNA was derived from a separate gene, CYP3A5P, most likely a pseudogene evolved from CYP3A5.

Construction of full-length cDNA library of white flower Salvia ...

African Journals Online (AJOL)

In order to screen and isolate secondary metabolite biosynthesis related gene, we construct a cDNA library of white flower Salvia miltiorrhiza bge. f.alba. High quality of total RNA was successfully isolated from roots of white flower S. miltiorrhiza using modified CTAB method. Double strand cDNA was cloned into pDNR-LIB ...

Molecular cloning of alpha-amylases from cotton boll weevil, Anthonomus grandis and structural relations to plant inhibitors: an approach to insect resistance.

Science.gov (United States)

Oliveira-Neto, Osmundo B; Batista, João A N; Rigden, Daniel J; Franco, Octávio L; Falcão, Rosana; Fragoso, Rodrigo R; Mello, Luciane V; dos Santos, Roseane C; Grossi-de-Sá, Maria F

2003-01-01

Anthonomus grandis, the cotton boll weevil, causes severe cotton crop losses in North and South America. Here we demonstrate the presence of starch in the cotton pollen grains and young ovules that are the main A. grandis food source. We further demonstrate the presence of alpha-amylase activity, an essential enzyme of carbohydrate metabolism for many crop pests, in A. grandis midgut. Two alpha-amylase cDNAs from A. grandis larvae were isolated using RT-PCR followed by 5' and 3' RACE techniques. These encode proteins with predicted molecular masses of 50.8 and 52.7kDa, respectively, which share 58% amino acid identity. Expression of both genes is induced upon feeding and concentrated in the midgut of adult insects. Several alpha-amylase inhibitors from plants were assayed against A. grandis alpha-amylases but, unexpectedly, only the BIII inhibitor from rye kernels proved highly effective, with inhibitors generally active against other insect amylases lacking effect. Structural modeling of Amylag1 and Amylag2 showed that different factors seem to be responsible for the lack of effect of 0.19 and alpha-AI1 inhibitors on A. grandis alpha-amylase activity. This work suggests that genetic engineering of cotton to express alpha-amylase inhibitors may offer a novel route to A. grandis resistance.

Molecular cloning of lupin leghemoglobin cDNA

DEFF Research Database (Denmark)

Konieczny, A; Jensen, E O; Marcker, K A

1987-01-01

Poly(A)+ RNA isolated from root nodules of yellow lupin (Lupinus luteus, var. Ventus) has been used as a template for the construction of a cDNA library. The ds cDNA was synthesized and inserted into the Hind III site of plasmid pBR 322 using synthetic Hind III linkers. Clones containing sequences...... specific for nodules were selected by differential colony hybridization using 32P-labeled cDNA synthesized either from nodule poly(A)+ RNA or from poly(A)+ RNA of uninfected root as probes. Among the recombinant plasmids, the cDNA gene for leghemoglobin was identified. The protein structure derived from...... its nucleotide sequence was consistent with known amino acid sequence of lupin Lb II. The cloned lupin Lb cDNA hybridized to poly(A)+ RNA from nodules only, which is in accordance with the general concept, that leghemoglobin is expressed exclusively in nodules. Udgivelsesdato: 1987-null...

Interspecific hybridization and inbreeding effect in seed from a Eucalyptus grandis x E. urophylla clonal orchard in Brazil

Directory of Open Access Journals (Sweden)

Campinhos Eduardo N.

1998-01-01

Full Text Available We used allozyme markers to estimate the amount of natural hybridization between Eucalyptus grandis and E. urophylla in a 7.4-hectare commercial hybrid-seed orchard planted in Espírito Santo, Brazil. This orchard was planted in 1982 using a honeycomb design, with each hexagonal plot containing one E. grandis tree surrounded by six E. urophylla trees. There were 267 replicated hexagonal plots in the orchard. Seeds were harvested from the E. grandis clone only. The multilocus outcrossing rate estimated for the E. grandis clone averaged 70.2%, ranging from 33.0 to 99.0% among individual trees. Contaminant pollination, inferred from progeny genotypes containing alleles not present in the seven parental clones, accounted for 14.4% of the hybrid seed. Contaminant pollen was attributed to neighboring eucalyptus stands isolated from the orchard by a 400-m wide belt of native forest. Inbred and hybrid progenies were identified by their allozyme genotypes and transplanted to the field. Field growth of inbred progeny was 30% lower than that of hybrid plants at two and three years of age.

Cloning of cDNA encoding steroid 11β-hydroxylase (P450c11)

International Nuclear Information System (INIS)

Chua, S.C.; Szabo, P.; Vitek, A.; Grzeschik, K.H.; John, M.; White, P.C.

1987-01-01

The authors have isolated bovine and human adrenal cDNA clones encoding the adrenal cytochrome P-450 specific for 11β-hydroxylation (P450c11). A bovine adrenal cDNA library constructed in the bacteriophage λ vector gt10 was probed with a previously isolated cDNA clone corresponding to part of the 3' untranslated region of the 4.2-kilobase (kb) mRNA encoding P450c11. Several clones with 3.2-kb cDNA inserts were isolated. Sequence analysis showed that they overlapped the original probe by 300 base pairs (bp). Combined cDNA and RNA sequence data demonstrated a continuous open reading frame of 1509 bases. P450c11 is predicted to contain 479 amino acid residues in the mature protein in addition to a 24-residue amino-terminal mitochondrial signal sequence. A bovine clone was used to isolate a homologous clone with a 3.5-kb insert from a human adrenal cDNA library. A region of 1100 bp was 81% homologous to 769 bp of the coding sequence of the bovine cDNA except for a 400-bp segment presumed to be an unprocessed intron. Hybridization of the human cDNA to DNA from a panel of human-rodent somatic cell hybrid lines and in situ hybridization to metaphase spreads of human chromosomes localized the gene to the middle of the long arm of chromosome 8. These data should be useful in developing reagents for heterozygote detection and prenatal diagnosis of 11β-hydroxylase deficiency, the second most frequent cause of congenital adrenal hyperplasia

Recombinant Cry1Ia protein is highly toxic to cotton boll weevil (Anthonomus grandis Boheman) and fall armyworm (Spodoptera frugiperda).

Science.gov (United States)

Martins, E S; Aguiar, R W D S; Martins, N F; Melatti, V M; Falcão, R; Gomes, A C M M; Ribeiro, B M; Monnerat, R G

2008-05-01

To evaluate the activity of cry1Ia gene against cotton pests, Spodoptera frugiperda and Anthonomus grandis. Had isolated and characterized a toxin gene from the Bacillus thuringiensis S1451 strain which have been previously shown to be toxic to S. frugiperda and A. grandis. The toxin gene (cry1Ia) was amplified by PCR, sequenced, and cloned into the genome of a baculovirus. The Cry1Ia protein was expressed in baculovirus infected insect cells, producing protein inclusions in infected cells. The Cry1Ia protein has used in bioassays against to S. frugiperda and A. grandis. Bioassays using the purified recombinant protein showed high toxicity to S. frugiperda and A. grandis larvae. Molecular modelling of the Cry1Ia protein translated from the DNA sequence obtained in this work, showed that this protein possibly posses a similar structure to the Cry3A protein. Ultrastructural analysis of midgut cells from A. grandis incubated with the Cry1Ia toxin, showed loss of microvilli integrity. The results indicate that the cry1Ia is a good candidate for the construction of transgenic plants resistant to these important cotton pests.

Isolation and characterization of human glycophorin A cDNA clones by a synthetic oligonucleotide approach: nucleotide sequence and mRNA structure

International Nuclear Information System (INIS)

Siebert, P.D.; Fukuda, M.

1986-01-01

In an effort to understand the relationships among and the regulation of human glycophorins, the authors have isolated and characterized several glycophorin A-specific cDNA clones obtained from a human erythroleukemic K562 cell cDNA library. This was accomplished by using mixed synthetic oligonucleotides, corresponding to various regions of the known amino acid sequence, to prime the synthesis of the cDNA as well as to screen the cDNA library. They also used synthetic oligonucleotides to sequence the largest of the glycophorin cDNAs. The nucleotide sequence obtained suggests the presence of a potential leader peptide, consistent with the membrane localization of this glycoprotein. Examination of the structure of glycophorin mRNA by blot hybridization revealed the existence of several electrophoretically distinct mRNAs numbering three or four, depending on the size of the glycophorin cDNA used as a hybridization probe. The smaller cDNA hybridized to three mRNAs of approximately 2.8, 1.7, and 1.0 kilobases. In contrast, the larger cDNA hybridized to an additional mRNA of approximately 0.6 kilobases. Further examination of the relationships between these multiple mRNAs by blot hybridization was conducted with the use of exact-sequence oligonucleotide probes constructed from various regions of the cDNA representing portions of the amino acid sequence of glycophorin A with or without known homology with glycophorin B. In total, the results obtained are consistent with the hypothesis that the three larger mRNAs represent glycophorin A gene transcripts and that the smallest (0.6 kilobase) mRNA may be specific for glycophorin B

Molecular cloning and mammalian expression of human beta 2-glycoprotein I cDNA

DEFF Research Database (Denmark)

Kristensen, Torsten; Schousboe, Inger; Boel, Espen

1991-01-01

Human β2-glycoprotein (β2gpI) cDNA was isolated from a liver cDNA library and sequenced. The cDNA encoded a 19-residue hydrophobic signal peptide followed by the mature β2gpI of 326 amino acid residues. In liver and in the hepatoma cell line HepG2 there are two mRNA species of about 1.4 and 4.3 kb......, respectively, hybridizing specifically with the β2gpI cDNA. Upon isoelectric focusing, recombinant β2gpI obtained from expression of β2gpI cDNA in baby hamster kidney cells showed the same pattern of bands as β2gpI isolated from plasma, and at least 5 polypeptides were visible...

Infectious Maize rayado fino virus from cloned cDNA

Science.gov (United States)

Maize rayado fino virus (MRFV) is the type member of the marafiviruses within the family Tymoviridae. A cDNA clone from which infectious RNA can be transcribed was produced from a US isolate of MRFV (MRFV-US). Infectivity of transcripts derived from cDNA clones was demonstrated by infection of mai...

[cDNA library construction from panicle meristem of finger millet].

Science.gov (United States)

Radchuk, V; Pirko, Ia V; Isaenkov, S V; Emets, A I; Blium, Ia B

2014-01-01

The protocol for production of full-size cDNA using SuperScript Full-Length cDNA Library Construction Kit II (Invitrogen) was tested and high quality cDNA library from meristematic tissue of finger millet panicle (Eleusine coracana (L.) Gaertn) was created. The titer of obtained cDNA library comprised 3.01 x 10(5) CFU/ml in avarage. In average the length of cDNA insertion consisted about 1070 base pairs, the effectivity of cDNA fragment insertions--99.5%. The selective sequencing of cDNA clones from created library was performed. The sequences of cDNA clones were identified with usage of BLAST-search. The results of cDNA library analysis and selective sequencing represents prove good functionality and full length character of inserted cDNA clones. Obtained cDNA library from meristematic tissue of finger millet panicle represents good and valuable source for isolation and identification of key genes regulating metabolism and meristematic development and for mining of new molecular markers to conduct out high quality genetic investigations and molecular breeding as well.

Isolation of a cDNA clone complementary to sequences for a 34-kilodalton protein which is a pp60v-src substrate.

OpenAIRE

Tomasiewicz, H G; Cook-Deegan, R; Chikaraishi, D M

1984-01-01

We have isolated a partial cDNA clone containing sequences complementary to a mRNA encoding a 34- to 36-kilodalton normal chicken cell protein which is a substrate for pp60v-src kinase activity. Using this 34-kilodalton cDNA clone as a probe, we determined that the size of the 34-kilodalton mRNA was 1,100 nucleotides and the level of the 34-kilodalton RNA was the same in various tissues of mature chickens but was significantly higher in chicken embryo fibroblast cells.

Isolation and sequence of cDNA encoding a cytochrome P-450 from an insecticide-resistant strain of the house fly, Musca domestica.

OpenAIRE

Feyereisen, R; Koener, J F; Farnsworth, D E; Nebert, D W

1989-01-01

A cDNA expression library from phenobarbital-treated house fly (Musca domestica) was screened with rabbit antisera directed against partially purified house fly cytochrome P-450. Two overlapping clones with insert lengths of 1.3 and 1.5 kilobases were isolated. The sequence of a 1629-base-pair (bp) cDNA was obtained, with an open reading frame (nucleotides 81-1610) encoding a P-450 protein of 509 residues (Mr = 58,738). The insect P-450 protein contains a hydrophobic NH2 terminus and a 22-res...

Functional cloning using pFB retroviral cDNA expression libraries.

Science.gov (United States)

Felts, Katherine A; Chen, Keith; Zaharee, Kim; Sundar, Latha; Limjoco, Jamie; Miller, Anna; Vaillancourt, Peter

2002-09-01

Retroviral cDNA expression libraries allow the efficient introduction of complex cDNA libraries into virtually any mitotic cell type for screening based on gene function. The cDNA copy number per cell can be easily controlled by adjusting the multiplicity of infection, thus cell populations may be generated in which >90% of infected cells contain one to three cDNAs. We describe the isolation of two known oncogenes and one cell-surface receptor from a human Burkitt's lymphoma (Daudi) cDNA library inserted into the high-titer retroviral vector pFB.

In vitro ectomycorrhiza formation by monokaryotic and dikaryotic isolates of Pisolithus microcarpus in Eucalyptus grandis Formação de ectomicorrizas in vitro por isolados monocarióticos e dicarióticos de Pisolithus microcarpus em Eucalyptus grandis

Directory of Open Access Journals (Sweden)

Maurício Dutra Costa

2010-06-01

Full Text Available The formation of ectomycorrhizas by monokaryotic and dikaryotic isolates of Pisolithus microcarpus (Cooke & Massee G. Cunn. in Eucalyptus grandis W. Hill ex Maid. was studied by in vitro synthesis in Petri dishes. The formation of ectomycorrhizas was observed for all strains tested. Ectomycorrhizas formed by the monokaryotic strains presented a sheath of hyphae around the roots and a Hartig net limited to the epidermis layer, typical of the angiosperm ectomycorrhizas. Colonization rates, a measure of the number of ectomycorrhizas in relation to the total number of lateral root tips, varied from 23 to 62%. Some monokaryotic strains stimulated the formation of lateral roots, promoting increases of up to 109% above the control. The presence of some of the isolates in the in vitro synthesis medium stimulated the production of thicker lateral root tips. The dimensions of the lateral roots tips and ectomycorrhizas varied from one isolate to the next, indicating a variation in their capacity to provoke morphological changes in the host plant roots. The dikaryotic strain M5M11 presented higher values for lateral root yield, number of ectomycorrhizas, and colonization percentage than the corresponding monokaryotic strains, M5 and M11. This indicated the possibility of selecting compatible performing monokaryotic isolates for the yield of superior dikaryotic strains. The set of monokaryotic strains tested varied greatly in their ability to colonize E. grandis roots and cause secondary metabolism-related morphological changes in roots, providing a wealth of model systems for the study of genetic, physiological, and morphogenetic processes involved in Pisolithus-Eucalyptus ectomycorrhiza formation.A formação de ectomicorrizas por isolados monocarióticos e dicarióticos de Pisolithus microcarpus (Cooke & Massee G. Cunn. em Eucalyptus grandis W. Hill ex Maid. foi estudada usando-se o método de síntese in vitro em placas de Petri. A formação de

cDNA encoding a polypeptide including a hevein sequence

Energy Technology Data Exchange (ETDEWEB)

Raikhel, N.V.; Broekaert, W.F.; Namhai Chua; Kush, A.

1993-02-16

A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1,018 nucleotides long and includes an open reading frame of 204 amino acids.

Isolation and characterization of full-length cDNA clones coding for cholinesterase from fetal human tissues

International Nuclear Information System (INIS)

Prody, C.A.; Zevin-Sonkin, D.; Gnatt, A.; Goldberg, O.; Soreq, H.

1987-01-01

To study the primary structure and regulation of human cholinesterases, oligodeoxynucleotide probes were prepared according to a consensus peptide sequence present in the active site of both human serum pseudocholinesterase and Torpedo electric organ true acetylcholinesterase. Using these probes, the authors isolated several cDNA clones from λgt10 libraries of fetal brain and liver origins. These include 2.4-kilobase cDNA clones that code for a polypeptide containing a putative signal peptide and the N-terminal, active site, and C-terminal peptides of human BtChoEase, suggesting that they code either for BtChoEase itself or for a very similar but distinct fetal form of cholinesterase. In RNA blots of poly(A) + RNA from the cholinesterase-producing fetal brain and liver, these cDNAs hybridized with a single 2.5-kilobase band. Blot hybridization to human genomic DNA revealed that these fetal BtChoEase cDNA clones hybridize with DNA fragments of the total length of 17.5 kilobases, and signal intensities indicated that these sequences are not present in many copies. Both the cDNA-encoded protein and its nucleotide sequence display striking homology to parallel sequences published for Torpedo AcChoEase. These finding demonstrate extensive homologies between the fetal BtChoEase encoded by these clones and other cholinesterases of various forms and species

Cloning and functional expression of a human pancreatic islet glucose-transporter cDNA

International Nuclear Information System (INIS)

Permutt, M.A.; Koranyi, L.; Keller, K.; Lacy, P.E.; Scharp, D.W.; Mueckler, M.

1989-01-01

Previous studies have suggested that pancreatic islet glucose transport is mediated by a high-K m , low-affinity facilitated transporter similar to that expressed in liver. To determine the relationship between islet and liver glucose transporters, liver-type glucose-transporter cDNA clones were isolated from a human liver cDNA library. The liver-type glucose-transporter cDNA clone hybridized to mRNA transcripts of the same size in human liver and pancreatic islet RNA. A cDNA library was prepared from purified human pancreatic islet tissue and screened with human liver-type glucose-transporter cDNA. The authors isolated two overlapping cDNA clones encompassing 2600 base pairs, which encode a pancreatic islet protein identical in sequence to that of the putative liver-type glucose-transporter protein. Xenopus oocytes injected with synthetic mRNA transcribed from a full-length cDNA construct exhibited increased uptake of 2-deoxyglucose, confirming the functional identity of the clone. These cDNA clones can now be used to study regulation of expression of the gene and to assess the role of inherited defects in this gene as a candidate for inherited susceptibility to non-insulin-dependent diabetes mellitus

Cloning of the human androgen receptor cDNA

International Nuclear Information System (INIS)

Govindan, M.V.; Burelle, M.; Cantin, C.; Kabrie, C.; Labrie, F.; Lachance, Y.; Leblanc, G.; Lefebvre, C.; Patel, P.; Simard, J.

1988-01-01

The authors discuss how in order to define the functional domains of the human androgen receptor, complementary DNA (cDNA) clones encoding the human androgen receptor (hAR) have been isolated from a human testis λgtll cDNA library using synthetic oligonnucleotide probes, homologous to segments of the human glucocorticoid, estradiol and progesterone receptors. The cDNA clones corresponding to the human glucocorticoid, estradiol and progesterone receptors were eliminated after cross-hybridization with their respective cDNA probes and/or after restriction mapping of the cDNA clones. The remaining cDNA clones were classified into different groups after analysis by restriction digestion and cross-hybridization. Two of the largest cDNA clones from each group were inserted into an expression vector in both orientations. The linearized plasmids were used as templates in in vitro transcription with T7 RNA polymerase. Subsequent in vitro translation of the purified transcripts in rabbit reticulocyte lysate followed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) permitted the characterization of the encoded polyeptides. The expressed proteins larger than 30,000 Da were analyzed for their ability to bind tritium-labelled dihydrotestosterone ([ 3 H] DHT) with high affinity and specificity

(+)-(10R)-Germacrene A synthase from goldenrod, Solidago canadensis; cDNA isolation, bacterial expression and functional analysis.

Science.gov (United States)

Prosser, Ian; Phillips, Andy L; Gittings, Simon; Lewis, Mervyn J; Hooper, Antony M; Pickett, John A; Beale, Michael H

2002-08-01

Profiling of sesquiterpene hydrocarbons in extracts of goldenrod, Solidago canadensis, by GC-MS revealed the presence of both enantiomers of germacrene D and lesser amounts of germacrene A, alpha-humulene, and beta-caryophyllene. A similarity-based cloning strategy using degenerate oligonucleotide primers, based on conserved amino acid sequences in known plant sesquiterpene synthases and RT-PCR, resulted in the isolation of a full length sesquiterpene synthase cDNA. Functional expression of the cDNA in E. coli, as an N-terminal thioredoxin fusion protein using the pET32b vector yielded an enzyme that was readily purified by nickel-chelate affinity chromatography. Chiral GC-MS analysis of products from of (3)H- and (2)H-labelled farnesyl diphosphate identified the enzyme as (+)-(10R)-germacrene A synthase. Sequence analysis and molecular modelling was used to compare this enzyme with the mechanistically related epi-aristolochene synthase from tobacco.

Isolation of cDNA clones coding for human tissue factor: primary structure of the protein and cDNA

International Nuclear Information System (INIS)

Spicer, E.K.; Horton, R.; Bloem, L.

1987-01-01

Tissue factor is a membrane-bound procoagulant protein that activates the extrinsic pathway of blood coagulation in the presence of factor VII and calcium. λ Phage containing the tissue factor gene were isolated from a human placental cDNA library. The amino acid sequence deduced from the nucleotide sequence of the cDNAs indicates that tissue factor is synthesized as a higher molecular weight precursor with a leader sequence of 32 amino acids, while the mature protein is a single polypeptide chain composed of 263 residues. The derived primary structure of tissue factor has been confirmed by comparison to protein and peptide sequence data. The sequence of the mature protein suggests that there are three distinct domains: extracellular, residues 1-219; hydrophobic, residues 220-242; and cytoplasmic, residues 243-263. Three potential N-linked carbohydrate attachment sites occur in the extracellular domain. The amino acid sequence of tissue factor shows no significant homology with the vitamin K-dependent serine proteases, coagulation cofactors, or any other protein in the National Biomedical Research Foundation sequence data bank (Washington, DC)

Micropropagation of Eucalyptus grandis and nitens using tissue culture techniques

Energy Technology Data Exchange (ETDEWEB)

Furze, M.J.; Creswell, C.F.

1985-01-01

Experiments with nodal explants of E. grandis and E. nitens seedlings and E. grandis coppice shoots showed that a large number of plants can be produced from a single explant using micropropagation. The percentage of micropropagated shoots that formed roots was about 90% for E. grandis and 80% for E. nitens. For both species, about 90% of the rooted shoots survived after hardening off. 9 references.

Cloning and expression of cDNA coding for bouganin.

Science.gov (United States)

den Hartog, Marcel T; Lubelli, Chiara; Boon, Louis; Heerkens, Sijmie; Ortiz Buijsse, Antonio P; de Boer, Mark; Stirpe, Fiorenzo

2002-03-01

Bouganin is a ribosome-inactivating protein that recently was isolated from Bougainvillea spectabilis Willd. In this work, the cloning and expression of the cDNA encoding for bouganin is described. From the cDNA, the amino-acid sequence was deduced, which correlated with the primary sequence data obtained by amino-acid sequencing on the native protein. Bouganin is synthesized as a pro-peptide consisting of 305 amino acids, the first 26 of which act as a leader signal while the 29 C-terminal amino acids are cleaved during processing of the molecule. The mature protein consists of 250 amino acids. Using the cDNA sequence encoding the mature protein of 250 amino acids, a recombinant protein was expressed, purified and characterized. The recombinant molecule had similar activity in a cell-free protein synthesis assay and had comparable toxicity on living cells as compared to the isolated native bouganin.

Actividad antianémica de la Cassia grandis L. The antianemic activity of Cassia grandis L.

Directory of Open Access Journals (Sweden)

Juana Tillán Capó

2004-12-01

Full Text Available La población de la zona oriental de Cuba refiere los efectos beneficiosos del uso tradicional de la Cassia grandis L. en la anemia, con la utilización del polvo seco obtenido del fruto como un suplemento nutricional. El objetivo fue evaluar dicho efecto en un modelo experimental de anemia ferropénica en ratas, inducido por sucesivas extracciones de sangre y administración de una dieta carente de hierro. Todos los animales fueron sometidos durante 15 días a una dieta semisintética deficiente en hierro y a extracciones de sangre 3 veces por semana hasta lograr concentraciones de hemoglobina en sangre menores de 9 g/dL. Se formaron 3 grupos y se mantuvo la misma dieta: grupo I sin suplementar, grupo II suplementado con 15 mg de hierro/kg de dieta y grupo III la misma cantidad de hierro más 750 mg/kg de peso corporal de polvo seco de Cassia grandis L. durante otros 15 días. Al finalizar se determinaron las concentraciones de hierro, hemoglobina y hematócrito en sangre. Las concentraciones medias de hemoglobina al cabo de los 15 días de tratamiento fueron significativamente diferentes en los 3 grupos experimentales, con resultados mayores en el grupo suplementado con hierro y Cassia grandis L.; también en este grupo se observó un incremento significativo de los valores medios de hierro en plasma en relación con los valores obtenidos en los animales no suplementados y en los animales que recibieron solamente hierro en la dieta. El porcentaje de hematócrito no mostró diferencia significativa entre tratamiento. Los resultados corroboran el uso popular y tradicional de la Cassia grandis L. en los estados anémicos, al mejorar la utilización del hiero y la producción de hemoglobina.The population of the eastern zone of Cuba refers to the benefitial effects of the traditional use of Cassia grandis L. in anemia by using dry powder obtained from the fruit as a nutritional supplement. The objective was to evaluate this effect in an

Estimativa do tempo de vaporização de toras de Eucalyptus grandis Steaming times estimates for Eucalyptus grandis logs

Directory of Open Access Journals (Sweden)

Fred Willians Calonego

2006-06-01

Full Text Available O objetivo deste estudo foi adequar o modelo geral de determinação do tempo de vaporização de toras, proposto por Steinhagen et al. (1980, para a madeira de Eucalyptus grandis. Para tanto, foram coletadas toras de 20 a The aim this study was to adjust the general model for determining log steaming time, proposed by Steinhagen et al. (1980, for Eucalyptus grandis wood. In order to do so, logs with diameter from 20 to <25, 25 to <30 and 30 to <35 cm were collected from 14 trees of Eucalyptus grandis derived from the `Horto Florestal' nursery, Manduri, São Paulo. A thermocouple was inserted into each log near its center. The logs were steamed during 20 hours at 90ºC and 100% relative humidity. A data logger recorded the temperatures during the thermal treatment. It was concluded that the Steinhagen et al. (1980 model cannot be directly used for this species in study and corrections factors are proposed for the utilization of the general model for log steaming time, developed by Steinhagen et al. (1980, for Eucalyptus grandis wood.

Wood Permeability in Eucalyptus grandis and Eucalyptus dunnii

Directory of Open Access Journals (Sweden)

Raphael Nogueira Rezende

2017-12-01

Full Text Available ABSTRACT The objective of this study was to evaluate the flow of air and water in Eucalyptus grandis and Eucalyptus dunnii wood. Wood was collected from four trees aged 37 years in an experimental plantation of the Federal University of Lavras, Brazil. Planks were cut off the basal logs to produce specimens for air and water permeability testing. Results indicated that the longitudinal permeability to air and water of E. grandis wood were, on average, 5% and 10% higher, respectively, than that of E. dunnii wood. E. grandis and E. dunnii wood showed neither air nor water flow in the test for permeability transversal to the fibers, and longitudinal permeability to air exceeded that to water by approximately 50 fold in both species.

Hea õpetamise grandi saajad

Index Scriptorium Estoniae

2016-01-01

Õpetamise uurimist alustavad grandi toel Andrus Org, Riina Oruaas, Natalja Zagura (humanitaar- ja kunstide valdkond), Stefano Braghiroli, Aet Kiisla, Tiiu Taur (sotsiaalteadused), Marje Oona, Oivi Uibo, Daisy Volmer (meditsiinitead.), Svetlana Ganina, Natalja Lepik ja Raivo Raid (loodus- ja täppistead.)

Natural and synthetic oviposition stimulants forCatolaccus grandis (Burks) females.

Science.gov (United States)

Guerra, A A; Martinez, S; Sonia Del Rio, H

1994-07-01

Oviposition behavior was elicited fromCatolaccus grandis (Burks) (Hymenoptera: Pteromalidae) females, an ectoparasitoid of the boll weevil,Anthonomus grandis Boheman (Coleoptera: Curculionidae), by smears of freshly cut cotton bolls or smears of extracts prepared with boll weevil damaged or undamaged cotton boll tissues. Oviposition behavior was also elicited fromC. grandis females by smears made withn-pentane,n-hexane,n-heptane, and isooctane. This is the first report of oviposition behavior elicited for any parasitoid by these short-chain saturated hydrocarbons (alkanes), introducing a new concept on the chemical mediation of parasitoid behavior during host selection. Oviposition behavior was also elicited fromC. grandis females by volatiles emanating from an artificial diet devoid of insect components that was specifically developed for the in vitro rearing of ectoparasitoids. The possible use of a synergistic combination ofn-hexane and diet to optimize the mechanized production of noncontaminated eggs is also discussed.

CDNA cloning, characterization and expression of an endosperm-specific barley peroxidase

DEFF Research Database (Denmark)

Rasmussen, Søren Kjærsgård; Welinder, K.G.; Hejgaard, J.

1991-01-01

A barley peroxidase (BP 1) of pI ca. 8.5 and M(r) 37000 has been purified from mature barley grains. Using antibodies towards peroxidase BP 1, a cDNA clone (pcR7) was isolated from cDNA expression library. The nucleotide sequence of pcR7 gave a derived amino acid sequence identical to the 158 C...

Cloning of the cDNA for human 12-lipoxygenase

International Nuclear Information System (INIS)

Izumi, T.; Hoshiko, S.; Radmark, O.; Samuelsson, B.

1990-01-01

A full-length cDNA clone encoding 12-lipoxygenase was isolated from a human platelet cDNA library by using a cDNA for human reticulocyte 15-lipoxygenase as probe for the initial screening. The cDNA had an open reading frame encoding 662 amino acid residues with a calculated molecular weight of 75,590. Three independent clones revealed minor heterogeneities in their DNA sequences. Thus, in three positions of the deduced amino acid sequence, there is a choice between two different amino acids. The deduced sequence from the clone plT3 showed 65% identity with human reticulocyte 15-lipoxygenase and 42% identity with human leukocyte 5-lipoxygenase. The 12-lipoxygenase cDNA recognized a 3.0-kilobase mRNA species in platelets and human erythroleukemia cells (HEL cells). Phorbol 12-tetradecanoyl 13-acetate induced megakaryocytic differentiation of HEL cells and 12-lipoxygenase activity and increased mRNA for 12-lipoxygenase. The identity of the cloned 12-lipoxygenase was assured by expression in a mammalian cell line (COS cells). Human platelet 12-lipoxygenase has been difficult to purify to homogeneity. The cloning of this cDNA will increase the possibilities to elucidate the structure and function of this enzyme

Prospects of Tectona Grandis as a Feedstock for Biodiesel

International Nuclear Information System (INIS)

Sarin, Amit; Singh, Meetu; Sharma, Neerja; Singh, N. P.

2017-01-01

The limited availability of fossil fuels has encouraged the need of replacement fuels of renewable nature. Among the renewable fuels, biodiesel produced from oil seeds and food wastes has been favored by the majority of researchers. In this study, Tectona Grandis seed oil has been investigated as a non-edible feedstock for biodiesel. The oil content of seed is 43% which makes it suitable for commercial production of biodiesel. The synthesis of biodiesel from T. Grandis oil was done with transesterification reaction giving high percentage yield of biodiesel which reached to 89%. The T. Grandis biodiesel was subjected to determine various physicochemical parameters by standard testing methods and found in agreement with the ASTM D-6751 and EN-14214 standards. The fatty-acid methyl ester composition for the biodiesel is composed of 42.71% oleic acid, 13.1% palmitic acid, and 31.51% linoleic acid. The biodiesel showed low oxidation stability which is attributed to high percentage of unsaturation. To address this issue, synthetic antioxidants were added to increase its resistance towards oxidation. By considering all the parameters, the present study reveals that T. Grandis seed oil is reliable for the production of biodiesel with encouraging probability in future.

Prospects of Tectona Grandis as a Feedstock for Biodiesel

Energy Technology Data Exchange (ETDEWEB)

Sarin, Amit, E-mail: amit.sarin@yahoo.com [Department of Physical Sciences, I.K. Gujral Punjab Technical University, Kapurthala (India); Singh, Meetu [Department of Applied Sciences, I.K. Gujral Punjab Technical University, Kapurthala (India); Sharma, Neerja [PG Department of Physics and Electronics, DAV College, Amritsar (India); Singh, N. P. [Department of Planning and External Development, I.K. Gujral Punjab Technical University, Kapurthala (India)

2017-10-26

The limited availability of fossil fuels has encouraged the need of replacement fuels of renewable nature. Among the renewable fuels, biodiesel produced from oil seeds and food wastes has been favored by the majority of researchers. In this study, Tectona Grandis seed oil has been investigated as a non-edible feedstock for biodiesel. The oil content of seed is 43% which makes it suitable for commercial production of biodiesel. The synthesis of biodiesel from T. Grandis oil was done with transesterification reaction giving high percentage yield of biodiesel which reached to 89%. The T. Grandis biodiesel was subjected to determine various physicochemical parameters by standard testing methods and found in agreement with the ASTM D-6751 and EN-14214 standards. The fatty-acid methyl ester composition for the biodiesel is composed of 42.71% oleic acid, 13.1% palmitic acid, and 31.51% linoleic acid. The biodiesel showed low oxidation stability which is attributed to high percentage of unsaturation. To address this issue, synthetic antioxidants were added to increase its resistance towards oxidation. By considering all the parameters, the present study reveals that T. Grandis seed oil is reliable for the production of biodiesel with encouraging probability in future.

Clonal differences in log end splitting in Eucalyptus grandis in ...

African Journals Online (AJOL)

This paper discusses the juvenile–mature correlation of log end splitting among Eucalyptus grandis clones from two trials and how differences in splitting relate to differences in wood density, pith-to-bark gradient and growth rate. Two approximately 20-year-old Eucalyptus grandis clonal trials at Bergvliet plantation were ...

Characterization of cDNA for PMT: a Partial Nicotine Biosynthesis-Related Gene Isolated from Indonesian Local Tobacco (Nicotiana tabacum cv. Sindoro1

Directory of Open Access Journals (Sweden)

SESANTI BASUKI

2013-12-01

Full Text Available Nicotine is the major alkaloid compound in cultivated tobacco (Nicotiana tabacum that could potentially be converted into carcinogenic compound (nor-nicotine. The PMT gene encoding putrescine N-methyltransferase (PMT is one of the two key genes that play a prominent role in nicotine biosynthesis. The aimed of this study was to isolate and characterize the cDNA sequence originated from Indonesian local tobacco cv. Sindoro1 (Ntpmt_Sindoro1. The results showed that the Ntpmt_Sindoro1 was 1124 bp in length. This cDNA fragment encodes for 374 amino acid residues. The predicted polypeptide from the cDNA is a hidrophilic protein, and has a predicted molecular weight of 40.95 kDa. The predicted amino acids sequence also showed high similarity to the PMT gene product Nicotiana sp. available in the GenBank data base. The amino acid sequences also exert conserved residues specifically exhibited only by PMT gene originated from N. tabacum. Clustering analysis revealed that Ntpmt_Sindoro1 belongs to the same clade as the PMT3 gene, a member of the N. tabacum PMT gene family. The Ntpmt_Sindoro1 cDNA sequence covering exon1-exon8 of the PMT gene fragment has been registered in the GenBank data base, under the accession number JX978277.

Sequence of a cloned cDNA encoding human ribosomal protein S11

Energy Technology Data Exchange (ETDEWEB)

Lott, J B; Mackie, G A

1988-02-11

The authors have isolated a cloned cDNA that encodes human ribosomal protein (rp) S11 by screening a human fibroblast cDNA library with a labelled 204 bp DNA fragment encompassing residues 212-416 of pRS11, a rat rp Sll cDNA clone. The human rp S11 cloned cDNA consists of 15 residues of the 5' leader, the entire coding sequence and all 51 residues of the 3' untranslated region. The predicted amino acid sequence of 158 residues is identical to rat rpS11. The nucleotide sequence in the coding region differs, however, from that in rat in the first position in two codons and in the third position in 44 codons.

cDNA encoding a polypeptide including a hevein sequence

Energy Technology Data Exchange (ETDEWEB)

Raikhel, Natasha V. (Okemos, MI); Broekaert, Willem F. (Dilbeek, BE); Chua, Nam-Hai (Scarsdale, NY); Kush, Anil (New York, NY)

1993-02-16

A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a pu GOVERNMENT RIGHTS This application was funded under Department of Energy Contract DE-AC02-76ER01338. The U.S. Government has certain rights under this application and any patent issuing thereon.

Growth and yield models for Eucalyptus grandis grown in Swaziland ...

African Journals Online (AJOL)

The aim of this study was to develop a stand-level growth and yield model for short-rotationEucalyptus grandis grown for pulp wood production at Piggs Peak in Swaziland. The data were derived from a Nelder 1a spacing trial established with E. grandis clonal cuttings in 1998 and terminated in 2005. Planting density ...

Molecular cloning and nucleotide sequence of cDNA for human liver arginase

International Nuclear Information System (INIS)

Haraguchi, Y.; Takiguchi, M.; Amaya, Y.; Kawamoto, S.; Matsuda, I.; Mori, M.

1987-01-01

Arginase (EC3.5.3.1) catalyzes the last step of the urea cycle in the liver of ureotelic animals. Inherited deficiency of the enzyme results in argininemia, an autosomal recessive disorder characterized by hyperammonemia. To facilitate investigation of the enzyme and gene structures and to elucidate the nature of the mutation in argininemia, the authors isolated cDNA clones for human liver arginase. Oligo(dT)-primed and random primer human liver cDNA libraries in λ gt11 were screened using isolated rat arginase cDNA as a probe. Two of the positive clones, designated λ hARG6 and λ hARG109, contained an overlapping cDNA sequence with an open reading frame encoding a polypeptide of 322 amino acid residues (predicted M/sub r/, 34,732), a 5'-untranslated sequence of 56 base pairs, a 3'-untranslated sequence of 423 base pairs, and a poly(A) segment. Arginase activity was detected in Escherichia coli cells transformed with the plasmid carrying λ hARG6 cDNA insert. RNA gel blot analysis of human liver RNA showed a single mRNA of 1.6 kilobases. The predicted amino acid sequence of human liver arginase is 87% and 41% identical with those of the rat liver and yeast enzymes, respectively. There are several highly conserved segments among the human, rat, and yeast enzymes

Long-Term Effects of Exotic Tree Species ( Tectona grandis Linn. F ...

African Journals Online (AJOL)

Long-Term Effects of Exotic Tree Species ( Tectona grandis Linn. F.) on the Status of Extractable Micronutrients in the ... The study therefore implied that Tectona grandis has an extractive property on micronutrient particularly on soils that are low in these nutrients. Nigerian Journal of Soil and Environmental Research Vol.

[Construction of forward and reverse subtracted cDNA libraries between muscle tissue of Meishan and Landrace pigs].

Science.gov (United States)

Xu, De-Quan; Zhang, Yi-Bing; Xiong, Yuan-Zhu; Gui, Jian-Fang; Jiang, Si-Wen; Su, Yu-Hong

2003-07-01

Using suppression subtractive hybridization (SSH) technique, forward and reverse subtracted cDNA libraries were constructed between Longissimus muscles from Meishan and Landrace pigs. A housekeeping gene, G3PDH, was used to estimate the efficiency of subtractive cDNA. In two cDNA libraries, G3PDH was subtracted very efficiently at appropriate 2(10) and 2(5) folds, respectively, indicating that some differentially expressed genes were also enriched at the same folds and the two subtractive cDNA libraries were very successful. A total of 709 and 673 positive clones were isolated from forward and reverse subtracted cDNA libraries, respectively. Analysis of PCR showed that most of all plasmids in the clones contained 150-750 bp inserts. The construction of subtractive cDNA libraries between muscle tissue from different pig breeds laid solid foundations for isolating and identifying the genes determining muscle growth and meat quality, which will be important to understand the mechanism of muscle growth, determination of meat quality and practice of molecular breeding.

Zeuxine grandis Seidenf. (Orchidaceae - an Addition to the Orchid Flora of India

Directory of Open Access Journals (Sweden)

Avishek Bhattacharjee

2010-09-01

Full Text Available Zeuxine grandis Seidenf., is being reported for the first time from India. We provide line drawing and color photo of specimen of Z. grandis in support of our treatment and to facilitate identification of the species.

Brain cDNA clone for human cholinesterase

International Nuclear Information System (INIS)

McTiernan, C.; Adkins, S.; Chatonnet, A.; Vaughan, T.A.; Bartels, C.F.; Kott, M.; Rosenberry, T.L.; La Du, B.N.; Lockridge, O.

1987-01-01

A cDNA library from human basal ganglia was screened with oligonucleotide probes corresponding to portions of the amino acid sequence of human serum cholinesterase. Five overlapping clones, representing 2.4 kilobases, were isolated. The sequenced cDNA contained 207 base pairs of coding sequence 5' to the amino terminus of the mature protein in which there were four ATG translation start sites in the same reading frame as the protein. Only the ATG coding for Met-(-28) lay within a favorable consensus sequence for functional initiators. There were 1722 base pairs of coding sequence corresponding to the protein found circulating in human serum. The amino acid sequence deduced from the cDNA exactly matched the 574 amino acid sequence of human serum cholinesterase, as previously determined by Edman degradation. Therefore, our clones represented cholinesterase rather than acetylcholinesterase. It was concluded that the amino acid sequences of cholinesterase from two different tissues, human brain and human serum, were identical. Hybridization of genomic DNA blots suggested that a single gene, or very few genes coded for cholinesterase

Isolation of a cDNA for a Growth Factor of Vascular Endothelial Cells from Human Lung Cancer Cells: Its Identity with Insulin‐like Growth Factor II

Science.gov (United States)

Hagiwara, Koichi; Kobayashi, Tatsuo; Tobita, Masato; Kikyo, Nobuaki; Yazaki, Yoshio

1995-01-01

We have found growth‐promoting activity for vascular endothelial cells in the conditioned medium of a human lung cancer cell line, T3M‐11. Purification and characterization of the growth‐promoting activity have been carried out using ammonium sulfate precipitation and gel‐exclusion chromatography. The activity migrated as a single peak just after ribonuclease. It did not bind to a heparin affinity column. These results suggest that the activity is not a heparin‐binding growth factor (including fibroblast growth factors) or a vascular endothelial growth factor. To identify the molecule exhibiting the growth‐promoting activity, a cDNA encoding the growth factor was isolated through functional expression cloning in COS‐1 cells from a cDNA library prepared from T3M‐11 cells. The nucleotide sequence encoded by the cDNA proved to be identical with that of insulin‐like growth factor II. PMID:7730145

[Habitat factor analysis for Torreya grandis cv. Merrillii based on spatial information technology].

Science.gov (United States)

Wang, Xiao-ming; Wang, Ke; Ao, Wei-jiu; Deng, Jin-song; Han, Ning; Zhu, Xiao-yun

2008-11-01

Torreya grandis cv. Merrillii, a tertiary survival plant, is a rare tree species of significant economic value and expands rapidly in China. Its special habitat factor analysis has the potential value to provide guide information for its planting, management, and sustainable development, because the suitable growth conditions for this tree species are special and strict. In this paper, the special habitat factors for T. grandis cv. Merrillii in its core region, i.e., in seven villages of Zhuji City, Zhejiang Province were analyzed with Principal Component Analysis (PCA) and a series of data, such as IKONOS image, Digital Elevation Model (DEM), and field survey data supported by the spatial information technology. The results showed that T. grandis cv. Merrillii exhibited high selectivity of environmental factors such as elevation, slope, and aspect. 96.22% of T. grandis cv. Merrillii trees were located at the elevation from 300 to 600 m, 97.52% of them were found to present on the areas whose slope was less than 300, and 74.43% of them distributed on sunny and half-sunny slopes. The results of PCA analysis indicated that the main environmental factors affecting the habitat of T. grandis cv. Merrillii were moisture, heat, and soil nutrients, and moisture might be one of the most important ecological factors for T. grandis cv. Merrillii due to the unique biological and ecological characteristics of the tree species.

The nucleotide sequence of human transition protein 1 cDNA

Energy Technology Data Exchange (ETDEWEB)

Luerssen, H; Hoyer-Fender, S; Engel, W [Universitaet Goettingen (West Germany)

1988-08-11

The authors have screened a human testis cDNA library with an oligonucleotide of 81 mer prepared according to a part of the published nucleotide sequence of the rat transition protein TP 1. They have isolated a cDNA clone with the length of 441 bp containing the coding region of 162 bp for human transition protein 1. There is about 84% homology in the coding region of the sequence compared to rat. The human cDNA-clone encodes a polypeptide of 54 amino acids of which 7 are different to that of rat.

Characterization of a human MSX-2 cDNA and its fragment isolated as a transformation suppressor gene against v-Ki-ras oncogene.

Science.gov (United States)

Takahashi, C; Akiyama, N; Matsuzaki, T; Takai, S; Kitayama, H; Noda, M

1996-05-16

A cDNA (termed CT124) encoding a carboxyl-terminal fragment of the human homeobox protein MSX-2 was found to induce flat reversion when expressed in v-Ki-ras-transformed NIH3T3 cells. Although the expression of endogenous MSX-2 gene is low in most of the normal adult tissues examined, it is frequently activated in carcinoma-derived cell lines. Likewise, the gene is inactive in NIH3T3 cells but is transcriptionally activated after transformation by v-Ki-ras oncogene, suggesting that the intact MSX-2 may play a positive, rather than suppressive, role in cell transformation. To test this possibility, we isolated a near full-length human MSX-2 cDNA and tested its activities in two cell systems, i.e. fibroblast and myoblast. In NIH3T3 fibroblasts, although the gene by itself failed to confer a transformed phenotype, antisense MSX-2 cDNA as well as truncated CT124 cDNA interfered with the transforming activities of v-Ki-ras oncogene. In C2C12 myoblasts, MSX-2 was found to suppress MyoD gene expression, as do activated ras oncogenes, under certain culture conditions, and CT124 was found to inhibit the activities of both MSX-2 and ras in this system as well. Our findings not only suggest that CT124 may act as a dominant suppressor of MSX-2 but also raise the possibility that MSX-2 gene may be an important downstream target for the Ras signaling pathways.

cDNA cloning of rat and human medium chain acyl-CoA dehydrogenase (MCAD)

International Nuclear Information System (INIS)

Matsubara, Y.; Kraus, J.P.; Rosenberg, L.E.; Tanaka, K.

1986-01-01

MCAD is one of three mitochondrial flavoenzymes which catalyze the first step in the β-oxidation of straight chain fatty acids. It is a tetramer with a subunit Mr of 45 kDa. MCAD is synthesized in the cytosol as a 49 kDa precursor polypeptide (pMCAD), imported into mitochondria, and cleaved to the mature form. Genetic deficiency of MCAD causes recurrent episodes of hypoglycemic coma accompanied by medium chain dicarboxylic aciduria. Employing a novel approach, the authors now report isolation of partial rat and human cDNA clones encoding pMCAD. mRNA encoding pMCAD was purified to near homogeneity by polysome immunoadsorption using polyclonal monospecific antibody. Single-stranded [ 32 P]labeled cDNA probe was synthesized using the enriched mRNA as template, and was used to screen directly 16,000 colonies from a total rat liver cDNA library constructed in pBR322. One clone (600 bp) was detected by in situ hybridization. Hybrid-selected translation with this cDNA yielded a 49 kDa polypeptide indistinguishable in size from rat pMCAD and immunoprecipitable with anti-MCAD antibody. Using the rat cDNA as probe, 43,000 colonies from a human liver cDNA library were screened. Four identical positive clones (400 bp) were isolated and positively identified by hybrid-selected translation and immunoprecipitation. The sizes of rat and human mRNAs encoding pMCAD were 2.2 kb and 2.4 kb, respectively, as determined by Northern blotting

Biology of Anastrepha grandis (Diptera: Tephritidae) in Different Cucurbits.

Science.gov (United States)

Bolzan, Anderson; Nava, Dori E; Garcia, Flávio R M; Valgas, Ricardo A; Smaniotto, Giovani

2015-06-01

Anastrepha grandis (Macquart) (Diptera: Tephritidae) is one of the main pests of cucurbits in Brazil. Losses occur due to the damage caused to the fruits and the embargo on exports, as A. grandis is considered a quarantine pest in countries that import Brazilian cucurbits. This study aimed to evaluate the development of A. grandis in hosts of the Cucurbitaceae family. The hosts used were stem squash (Cucurbita pepo L.), squash (Cucurbita moschata Duchesne), chayote [Sechium edule (Jacq.) Swartz], mini watermelon [Citrullus lanatus (Thunb.) Matsum & Nakai], Spanish melon (Cucumis melo L.), hybrid squash "Tetsukabuto" (C. moschata×Cucurbita maxima Duchesne), and salad cucumber (Cucumis sativus L.). We evaluated the viability and duration of egg-to-pupa period, pupal weight, sex ratio, and average number of pupae per fruit under controlled conditions of temperature, relative humidity, and photophase. The preoviposition and oviposition periods, fecundity, fertility, and longevity of females were determined for adults. Hosts of the genus Cucurbita provided a better development of A. grandis in comparison with other hosts, and presented a greater number of insects on fruit as well as higher infestation rate. Fecundity and longevity were also higher for females that developed in hosts of the genus Cucurbita, although values of these biological parameters varied between stem squash, squash, hybrid squash "Tetsukabuto." © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Cloning a cDNA for the lysosomal alpha-glucosidase

NARCIS (Netherlands)

KONINGS, A.; HUPKES, P.; Versteeg, R.; Grosveld, G.; Reuser, A.; Galjaard, H.

1984-01-01

Messenger RNA was isolated from monkey testes and size-fractionated on sucrose gradients. In vitro translation of these mRNA fractions resulted in nascent, labeled alpha-glucosidase that could be precipitated with anti human alpha-glucosidase antiserum. A cDNA library was constructed from the most

cDNA sequences of two inducible T-cell genes

Energy Technology Data Exchange (ETDEWEB)

Kwon, B.S. (Indiana Univ. School of Medicine, Indianapolis (USA) Guthrie Research Institute, Sayre, PA (USA)); Weissman, S.M. (Yale Univ., New Haven, CT (USA))

1989-03-01

The authors have previously described a set of human T-lymphocyte-specific cDNA clones isolated by a modified differential screening procedure. Apparent full-length cDNAs containing the sequences of 14 of the 16 initial isolates were sequenced and were found to represent five different species of mRNA; three of the five species were identical to previously reported cDNA sequences of preproenkephalin, T-cell-replacing factor, and a serine esterase, respectively. The other two species, 4-1BB and L2G25B, were inducible sequences found in mRNA from both a cytolytic T-lymphocyte and a helper T-lymphocyte clone and were not previously described in T-cell mRNA; these mRNA sequences encode peptides of 256 and 92 amino acids, respectively. Both peptides contain putative leader sequences. The protein encoded by 4-1BB also has a potential membrane anchor segment and other features also seen in known receptor proteins.

Molecular characterization of a Leishmania donovani cDNA clone with similarity to human 20S proteasome a-type subunit

DEFF Research Database (Denmark)

Christensen, C B; Jørgensen, L; Jensen, A T

2000-01-01

Using plasma from patients infected or previously infected with Leishmania donovanii, we isolated a L. donovanii cDNA clone with similarity to the proteasome a-type subunit from humans and other eukaryotes. The cDNA clone, designated LePa, was DNA sequenced and Northern blot analysis of L....... donovanii poly(A(+))mRNA indicated the isolation of a full length cDNA clone with a transcript size of 1.9 kb. The expressed recombinant LePa fusion protein induced proliferation of peripheral blood mononuclear cells in one out of seven patients who had suffered from visceral leishmaniasis. Plasma from 16...

Cloning and analysis of the mouse Fanconi anemia group A cDNA and an overlapping penta zinc finger cDNA.

Science.gov (United States)

Wong, J C; Alon, N; Norga, K; Kruyt, F A; Youssoufian, H; Buchwald, M

2000-08-01

Despite the cloning of four disease-associated genes for Fanconi anemia (FA), the molecular pathogenesis of FA remains largely unknown. To study FA complementation group A using the mouse as a model system, we cloned and characterized the mouse homolog of the human FANCA cDNA. The mouse cDNA (Fanca) encodes a 161-kDa protein that shares 65% amino acid sequence identity with human FANCA. Fanca is located at the distal region of mouse chromosome 8 and has a ubiquitous pattern of expression in embryonic and adult tissues. Expression of the mouse cDNA in human FA-A cells restores the cellular drug sensitivity to normal levels. Thus, the expression pattern, protein structure, chromosomal location, and function of FANCA are conserved in the mouse. We also isolated a novel zinc finger protein, Zfp276, which has five C(2)H(2) domains. Interestingly, Zfp276 is situated in the Fanca locus, and the 3'UTR of its cDNA overlaps with the last four exons of Fanca in a tail-to-tail manner. Zfp276 is expressed in the same tissues as Fanca, but does not complement the mitomycin C (MMC)-sensitive phenotype of FA-A cells. The overlapping genomic organization between Zfp276 and Fanca may have relevance to the disease phenotype of FA. Copyright 2000 Academic Press.

Description and Phylogeny of Urostyla grandis wiackowskii subsp. nov. (Ciliophora, Hypotricha) from an Estuarine Mangrove in Brazil.

Science.gov (United States)

Paiva, Thiago da Silva; Shao, Chen; Fernandes, Noemi Mendes; Borges, Bárbara do Nascimento; da Silva-Neto, Inácio Domingos

2016-01-01

Interphase specimens, aspects of physiological reorganization and divisional morphogenesis were investigated in a strain of a hypotrichous ciliate highly similar to Urostyla grandis Ehrenberg, (type species of Urostyla), collected from a mangrove area in the estuary of the Paraíba do Sul river (Rio de Janeiro, Brazil). The results revealed that albeit interphase specimens match with the known morphologic variability in U. grandis, morphogenetic processes have conspicuous differences. Parental adoral zone is entirely renewed during morphogenesis, and marginal cirri exhibit a unique combination of developmental modes, in which left marginal rows originate from multiple anlagen arising from innermost left marginal cirral row, whereas right marginal ciliature originates from individual within-row anlagen. Based on such characteristics, a new subspecies, namely U. grandis wiackowskii subsp. nov. is proposed, and consequently, U. grandis grandis Ehrenberg, stat. nov. is established. Bayesian and maximum-likelihood analyses of the 18S rDNA unambiguously placed U. grandis wiackowskii as adelphotaxon of a cluster formed by other U. grandis sequences. The implications of such findings to the systematics of Urostyla are discussed. © 2015 The Author(s) Journal of Eukaryotic Microbiology © 2015 International Society of Protistologists.

Cloning and sequence analysis of cDNA coding for rat nucleolar protein C23

International Nuclear Information System (INIS)

Ghaffari, S.H.; Olson, M.O.J.

1986-01-01

Using synthetic oligonucleotides as primers and probes, the authors have isolated and sequenced cDNA clones encoding protein C23, a putative nucleolus organizer protein. Poly(A + ) RNA was isolated from rat Novikoff hepatoma cells and enriched in C23 mRNA by sucrose density gradient ultracentrifugation. Two deoxyoligonuleotides, a 48- and a 27-mer, were synthesized on the basis of amino acid sequence from the C-terminal half of protein C23 and cDNA sequence data from CHO cell protein. The 48-mer was used a primer for synthesis of cDNA which was then inserted into plasmid pUC9. Transformed bacterial colonies were screened by hybridization with 32 P labeled 27-mer. Two clones among 5000 gave a strong positive signal. Plasmid DNAs from these clones were purified and characterized by blotting and nucleotide sequence analysis. The length of C23 mRNA was estimated to be 3200 bases in a northern blot analysis. The sequence of a 267 b.p. insert shows high homology with the CHO cDNA with only 9 nucleotide differences and an identical amino acid sequence. These studies indicate that this region of the protein is highly conserved

Styl RFLP recognized by a human IRBP cDNA localized to chromosome 10

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Chin, K S; Mathew, C G.P.; Fong, S L; Bridges, C D; Ponder, B A.J.

1988-02-25

A 2184 bp cDNA (H.4 IRBP) encoding human interstitial retinol-biding protein isolated from a human retina cDNA library in lambdagt10 by screening with a bovine IRBP cDNA probe. Styl identifies a 2-allele polymorphism with bands at 2.3 kb (Cl) and 1.95 kb (C2) and invariant bands at 1.1, 1.0 and 0.8kb. Codominant segregation was observed in two informative families. The RFLP was mapped to chromosome 10 using somatic cell hybrids. In situ hybridization suggests regional assignments near p11.2 -q11.2 with a secondary site of hybridization at q24-25.

RICD: A rice indica cDNA database resource for rice functional genomics

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Zhang Qifa

2008-11-01

Full Text Available Abstract Background The Oryza sativa L. indica subspecies is the most widely cultivated rice. During the last few years, we have collected over 20,000 putative full-length cDNAs and over 40,000 ESTs isolated from various cDNA libraries of two indica varieties Guangluai 4 and Minghui 63. A database of the rice indica cDNAs was therefore built to provide a comprehensive web data source for searching and retrieving the indica cDNA clones. Results Rice Indica cDNA Database (RICD is an online MySQL-PHP driven database with a user-friendly web interface. It allows investigators to query the cDNA clones by keyword, genome position, nucleotide or protein sequence, and putative function. It also provides a series of information, including sequences, protein domain annotations, similarity search results, SNPs and InDels information, and hyperlinks to gene annotation in both The Rice Annotation Project Database (RAP-DB and The TIGR Rice Genome Annotation Resource, expression atlas in RiceGE and variation report in Gramene of each cDNA. Conclusion The online rice indica cDNA database provides cDNA resource with comprehensive information to researchers for functional analysis of indica subspecies and for comparative genomics. The RICD database is available through our website http://www.ncgr.ac.cn/ricd.

Cloning and characterization of the human colipase cDNA

International Nuclear Information System (INIS)

Lowe, M.E.; Rosenblum, J.L.; McEwen, P.; Strauss, A.W.

1990-01-01

Pancreatic lipase hydrolyzes dietary triglycerides to monoglycerides and fatty acids. In the presence of bile salts, the activity of pancreatic lipase is markedly decreased. The activity can be restored by the addition of colipase, a low molecular weight protein secreted by the pancreas. The action of pancreatic lipase in the gut lumen is dependent upon its interaction with colipase. As a first step in elucidating the molecular events governing the interaction of lipase and colipase with each other and with fatty acids, a cDNA encoding human colipase was isolated from a λgt11 cDNA library with a rabbit polyclonal anti-human colipase antibody. The full-length 525 bp cDNA contained an open reading frame encoding 112 amino acids, including a 17 amino acid signal peptide. The predicted sequence contains 100% of the published protein sequence for human colipase determined by chemical methods, but predicts the presence of five additional NH 2 -terminal amino acids and four additional COOH-terminal amino acids. Comparison of the predicted protein sequence with the known sequences of colipase from other species reveals regions of extensive identity. The authors report, for the first time, a cDNA for colipase. The cDNA predicts a human procolipase an suggests that there may also be processing at the COOH-terminus. The regions of identity with colipase from other species will aid in defining the interaction with lipase and lipids through site-specific mutagenesis

High-resolution genetic maps of Eucalyptus improve Eucalyptus grandis genome assembly.

Science.gov (United States)

Bartholomé, Jérôme; Mandrou, Eric; Mabiala, André; Jenkins, Jerry; Nabihoudine, Ibouniyamine; Klopp, Christophe; Schmutz, Jeremy; Plomion, Christophe; Gion, Jean-Marc

2015-06-01

Genetic maps are key tools in genetic research as they constitute the framework for many applications, such as quantitative trait locus analysis, and support the assembly of genome sequences. The resequencing of the two parents of a cross between Eucalyptus urophylla and Eucalyptus grandis was used to design a single nucleotide polymorphism (SNP) array of 6000 markers evenly distributed along the E. grandis genome. The genotyping of 1025 offspring enabled the construction of two high-resolution genetic maps containing 1832 and 1773 markers with an average marker interval of 0.45 and 0.5 cM for E. grandis and E. urophylla, respectively. The comparison between genetic maps and the reference genome highlighted 85% of collinear regions. A total of 43 noncollinear regions and 13 nonsynthetic regions were detected and corrected in the new genome assembly. This improved version contains 4943 scaffolds totalling 691.3 Mb of which 88.6% were captured by the 11 chromosomes. The mapping data were also used to investigate the effect of population size and number of markers on linkage mapping accuracy. This study provides the most reliable linkage maps for Eucalyptus and version 2.0 of the E. grandis genome. © 2014 CIRAD. New Phytologist © 2014 New Phytologist Trust.

Eucalyptus grandis AND Eucalyptus dunnii USE FOR WOOD-CEMENT PANELS MANUFACTURING

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Setsuo Iwakiri

2008-03-01

Full Text Available This research evaluated the potential use of Eucalyptus grandis and Eucalyptus dunnii wood for wood-cement panelsmanufacturing. The boards were manufactured at the density of 1,20 g/cm³, using portland cement as mineral bonding and woodfurnish without treatment, treated in cold water and hot water. The wood furnish of Pinus taeda was used as control. The resultsindicated that it is not necessary to treat E. grandis and E. dunni wood for wood-cement board manufacturing. In relation to woodspecies, the board manufactured with E. dunnii showed lower values of mechanical properties. However, boards manufactured of E.grandis wood showed satisfactory results in comparison to boards of P. taeda and the referenced values of BISON process and otherproducts cited in the pertnent literature, indicating the high potential for wood-cement board manufacture of this tree species.

Purification of Single-Stranded cDNA Based on RNA Degradation Treatment and Adsorption Chromatography.

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Trujillo-Esquivel, Elías; Franco, Bernardo; Flores-Martínez, Alberto; Ponce-Noyola, Patricia; Mora-Montes, Héctor M

2016-08-02

Analysis of gene expression is a common research tool to study networks controlling gene expression, the role of genes with unknown function, and environmentally induced responses of organisms. Most of the analytical tools used to analyze gene expression rely on accurate cDNA synthesis and quantification to obtain reproducible and quantifiable results. Thus far, most commercial kits for isolation and purification of cDNA target double-stranded molecules, which do not accurately represent the abundance of transcripts. In the present report, we provide a simple and fast method to purify single-stranded cDNA, exhibiting high purity and yield. This method is based on the treatment with RNase H and RNase A after cDNA synthesis, followed by separation in silica spin-columns and ethanol precipitation. In addition, our method avoids the use of DNase I to eliminate genomic DNA from RNA preparations, which improves cDNA yield. As a case report, our method proved to be useful in the purification of single-stranded cDNA from the pathogenic fungus Sporothrix schenckii.

LEDGF/p75 Deficiency Increases Deletions at the HIV-1 cDNA Ends.

Science.gov (United States)

Bueno, Murilo T D; Reyes, Daniel; Llano, Manuel

2017-09-15

Processing of unintegrated linear HIV-1 cDNA by the host DNA repair system results in its degradation and/or circularization. As a consequence, deficient viral cDNA integration generally leads to an increase in the levels of HIV-1 cDNA circles containing one or two long terminal repeats (LTRs). Intriguingly, impaired HIV-1 integration in LEDGF/p75-deficient cells does not result in a correspondent increase in viral cDNA circles. We postulate that increased degradation of unintegrated linear viral cDNA in cells lacking the lens epithelium-derived growth factor (LEDGF/p75) account for this inconsistency. To evaluate this hypothesis, we characterized the nucleotide sequence spanning 2-LTR junctions isolated from LEDGF/p75-deficient and control cells. LEDGF/p75 deficiency resulted in a significant increase in the frequency of 2-LTRs harboring large deletions. Of note, these deletions were dependent on the 3' processing activity of integrase and were not originated by aberrant reverse transcription. Our findings suggest a novel role of LEDGF/p75 in protecting the unintegrated 3' processed linear HIV-1 cDNA from exonucleolytic degradation.

Boll weevil (Anthonomus grandis) population genomics as a tool for monitoring and management

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Despite the success of eradication efforts across most of the cotton-producing regions of the U.S., the cotton boll weevil (Anthonomus grandis grandis Boheman) remains a major pest of cotton in much of the New World. The area along the Texas border with northern Mexico has been a particularly troub...

Competition between Catolaccus grandis (Hymenoptera: Pteromalidae and Bracon vulgaris (Hymenoptera: Braconidae, parasitoids of the Boll Weevil

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Francisco de Sousa Ramalho

2007-05-01

Full Text Available The competition between populations of the parasitoids C. grandis and B. vulgaris was studied using larvae of Euscepes postfasciatus (Fairmaire as an alternative host. A series of biological parameters was observed and related to the competitive abilities of both parasitoid species. They were capable of colonizing and maintaining their populations regardless of host location. The population growth of C. grandis and B. vulgaris, based on fecundity was not affected by the competition. The parasitism and survivorship to the adult stage were affected by competition, except when the host was located at the bottom of the rearing cage. C. grandis performed better than B. vulgaris independently of the competition and host location, but it did not exclude the other species.Catolaccus grandis (Burks e Bracon vulgaris Ashmead são os principais parasitóides do bicudo-do-algodoeiro Anthonomus grandis Boheman no Nordeste do Brasil. É importante que se determinem as interações entre esses parasitóides e o seu efeito em programas de controle biológico dessa praga com os mesmos. A competição entre os parasitóides C. grandis e B. vulgaris foi estudada, utilizando-se larvas de Euscepes postfasciatus Fairmaire como hospedeiro alternativo. A fecundidade de C. grandis e B. vulgaris baseada na produção de ovos, não foi afetada pela competição, mas o parasitismo e a produção de adultos desses parasitóides foram afetados pela competição, exceto quando o hospedeiro se encontrava na base da caixa de criação. Independentemente da competição e da localização do hospedeiro, C. grandis apresentou melhor desempenho que B. vulgaris, mas não excluiu as populações da outra espécie de parasitóide.

Sanitary and nutritional characterization of honeybee colonies in Eucalyptus grandis plantations

OpenAIRE

Invernizzi, C.; Santos, E.; García, E.; Daners, G.; Di Landro, R.; Saadoun, A.; Cabrera, C.

2011-01-01

In Uruguay, many beekeepers transport their colonies to Eucalyptus grandis plantations at the end of the summer and autumn, obtaining important honey harvests. However, at the end of the flowering period the colonies become extremely weakened undergoing high levels of mortality. Nutritional and health problems could explain the weakening of colonies. In order to find out the causes for this weakening, colonies of the same size were taken to an E. grandis plantation, split up in three groups d...

CDNA encoding a polypeptide including a hevein sequence

Science.gov (United States)

Raikhel, Natasha V.; Broekaert, Willem F.; Chua, Nam-Hai; Kush, Anil

1995-03-21

A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the protein of the present invention and maltose binding protein produced by the E. coli.

Cloning and characterization of a cDNA encoding topoisomerase II in pea and analysis of its expression in relation to cell proliferation.

Science.gov (United States)

Reddy, M K; Nair, S; Tewari, K K; Mudgil, Y; Yadav, B S; Sopory, S K

1999-09-01

We have isolated and sequenced four overlapping cDNA clones to identify the full-length cDNA for topoisomerase II (PsTopII) from pea. Using degenerate primers, based on the conserved amino acid sequences of other eukaryotic type II topoisomerases, a 680 bp fragment was PCR-amplified with pea cDNA as template. This fragment was used as a probe to screen an oligo-dT-primed pea cDNA library. A partial cDNA clone was isolated that was truncated at the 3' end. RACE-PCR was employed to isolate the remaining portion of the gene. The total size of PsTopII is 4639 bp with an open reading frame of 4392 bp. The deduced amino acid sequence shows a strong homology to other eukaryotic topoisomerase II (topo II) at the N-terminus end. The topo II transcript was abundant in proliferative tissues. We also show that the level of topo II transcripts could be stimulated by exogenous application of growth factors that induced proliferation in vitro cultures. Light irradiation to etiolated tissue strongly stimulated the expression of topo II. These results suggest that topo II gene expression is up-regulated in response to light and hormones and correlates with cell proliferation. Besides, we have also isolated and analysed the 5'-flanking region of the pea TopII gene. This is first report on the isolation of a putative promoter for topoisomerase II from plants.

Multiplicação in vitro de Eucalyptus grandis sob estímulo com benzilaminopurina In vitro multiplication of Eucalyptus grandis under BAP pulse

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Wirifran Fernandes de Andrade

2006-12-01

Full Text Available O objetivo deste trabalho foi avaliar os efeitos do estímulo com benzilaminopurina (BAP na multiplicação in vitro de Eucalyptus grandis. Foram avaliadas as interações entre concentrações de BAP (0, 200, 400 e 600 mg L-1, tempo de exposição (1, 2 e 3 horas, pH (3 e 5,8 da solução e alterações morfológicas dos explantes. Semanalmente, foi determinada a massa da matéria fresca dos explantes. Aos 21 dias de cultura, foram avaliados: o número de brotações por tratamento, o número de brotações obtidas por explante (taxa de multiplicação, e foi realizado o resgate das plântulas para avaliação histológica. O pH não apresentou interação com os demais fatores estudados. Os tratamentos com BAP a 200 mg L-1, durante 1 e 2 horas, apresentaram-se como os mais indicados na multiplicação do E. grandis. Houve intensificação da divisão celular, no parênquima cortical e no procâmbio, representada pelo surgimento de meristemóides, em resposta aos tratamentos com 200 mg L-1 de BAP, durante 1 e 2 horas. O estímulo com BAP na multiplicação in vitro de E. grandis é viável.The objective of this work was to evaluate the effect of pulse on in vitro multiplication of Eucalyptus grandis. Interactions were evaluated among BAP concentrations (0, 200, 400 and 600 mg L-1, exposure time (1, 2 and 3 hours, pH values (3 and 5.8, and explant morphological changes. Fresh weight of the explants was determined weekly. At 21 days of culture, valuations were made of the number of shootings per treatment, number of shootings obtained per explant (multiplication rate, and seedlings rescue was accomplished for histological analysis. The pH values did not present any interaction with the other factors. The most significant treatments on E. grandis shoot multiplications were 200 mg L-1 of 6-BAP during 1 and 2 hours. There was an intensification of cell division in the cortical parenchyma and procambium, represented by the arising of meristems in

Construction of a T7 Human Lung Cancer cDNA Library

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Wentao YUE

2008-10-01

Full Text Available Background and objective Currently, only a limited numbers of tumor markers for non small lung cancer (NSCLC diagnosis, new biomarker, such as serum autoantibody may improve the early detection of lung cancer. Our objective is construction human lung squamous carcinoma and adenocarcinoma T7 phage display cDNA library from the tissues of NSCLC patients. Methods mRNA was isolated from a pool of total RNA extract from NSCLC tissues obtained from 5 adenocarcinomas and 5 squamous carcinomas, and then mRNA was reverse transcribed into double stranded cDNA. After digestion, the cDNA was inserted into T7Select 10-3 vector. The phage display cDNA library was constructed by package reaction in vitro and plate proliferation. Plaque assay and PCR were used to evaluate the library.Results Two T7 phage display cDNA library were established. Plaque assay show the titer of lung squamas carcinoma library was 1.8×106 pfu, and the adenocarcinoma library was 5×106 pfu. The phage titer of the amplified library were 3.2×1010 pfu/mL and 2.5×1010 pfu/mL. PCR amplification of random plaque show insert ratio were 100% (24/24 in adenocarcinoma library and 95.8% in human lung squamas carcinoma library (23/24. Insert range from 300 bp to 1 500 bp. Conclusion Two phage display cDNA library from NSCLC were constructed.

Cloning, sequencing, and expression of cDNA for human β-glucuronidase

International Nuclear Information System (INIS)

Oshima, A.; Kyle, J.W.; Miller, R.D.

1987-01-01

The authors report here the cDNA sequence for human placental β-glucuronidase (β-D-glucuronoside glucuronosohydrolase, EC 3.2.1.31) and demonstrate expression of the human enzyme in transfected COS cells. They also sequenced a partial cDNA clone from human fibroblasts that contained a 153-base-pair deletion within the coding sequence and found a second type of cDNA clone from placenta that contained the same deletion. Nuclease S1 mapping studies demonstrated two types of mRNAs in human placenta that corresponded to the two types of cDNA clones isolated. The NH 2 -terminal amino acid sequence determined for human spleen β-glucuronidase agreed with that inferred from the DNA sequence of the two placental clones, beginning at amino acid 23, suggesting a cleaved signal sequence of 22 amino acids. When transfected into COS cells, plasmids containing either placental clone expressed an immunoprecipitable protein that contained N-linked oligosaccharides as evidenced by sensitivity to endoglycosidase F. However, only transfection with the clone containing the 153-base-pair segment led to expression of human β-glucuronidase activity. These studies provide the sequence for the full-length cDNA for human β-glucuronidase, demonstrate the existence of two populations of mRNA for β-glucuronidase in human placenta, only one of which specifies a catalytically active enzyme, and illustrate the importance of expression studies in verifying that a cDNA is functionally full-length

Human pro. cap alpha. 1)(I) collagen: cDNA sequence for the C-propeptide domain

Energy Technology Data Exchange (ETDEWEB)

Maekelae, J K; Raassina, M; Virta, A; Vuorio, E

1988-01-11

The authors have previously constructed a cDNA clone pHCAL1, covering most of the C-terminal propeptide domain of human pro..cap alpha..1(I) collagen mRNA,by inserting a 678 bp EcoRI-XhoI fragment of cDNA into pBR322. Since the XhoI/SalI ligation prevented removal of the insert, they used the same strategy to obtain a similar clone in pUC8. RNA was isolated from fetal calvarial bones. The cDNA was digested with EcoRI and XhoI and fractionated on a 1 % agarose gel. Fragments of 650-700 bp were cloned in pUC8 at the polylinker site, which now permits easy removal of the insert. The new clone was named pHCAL1U since the RNA was isolated from another individual. The approach outlined is useful for studies on individual variation which is important to recognize when searching for disease-related mutations in type I collagen.

Construction and characterization of a full-length infectious cDNA clone of foot-and-mouth disease virus strain O/JPN/2010 isolated in Japan in 2010.

Science.gov (United States)

Nishi, Tatsuya; Onozato, Hiroyuki; Ohashi, Seiichi; Fukai, Katsuhiko; Yamada, Manabu; Morioka, Kazuki; Kanno, Toru

2016-06-01

A full-length infectious cDNA clone of the genome of a foot-and-mouth disease virus isolated from the 2010 epidemic in Japan was constructed and designated pSVL-f02. Transfection of Cos-7 or IBRS-2 cells with this clone allowed the recovery of infectious virus. The recovered virus had the same in vitro characterization as the parental virus with regard to antigenicity in neutralization and indirect immunofluorescence tests, plaque size and one-step growth. Pigs were experimentally infected with the parental virus or the recombinant virus recovered from pSVL-f02 transfected cells. There were no significant differences in clinical signs or antibody responses between the two groups, and virus isolation and viral RNA detection from clinical samples were similar. Virus recovered from transfected cells therefore retained the in vitro characteristics and the in vivo pathogenicity of their parental strain. This cDNA clone should be a valuable tool to analyze determinants of pathogenicity and mechanisms of virus replication, and to develop genetically engineered vaccines against foot-and-mouth disease virus. Copyright © 2016 Elsevier Ltd. All rights reserved.

Paramyosin from the parasitic mite Sarcoptes scabiei: cDNA cloning and heterologous expression.

Science.gov (United States)

Mattsson, J G; Ljunggren, E L; Bergström, K

2001-05-01

The burrowing mite Sarcoptes scabiei is the causative agent of the highly contagious disease sarcoptic mange or scabies. So far, there is no in vitro propagation system for S. scabiei available, and mites used for various purposes must be isolated from infected hosts. Lack of parasite-derived material has limited the possibilities to study several aspects of scabies, including pathogenesis and immunity. It has also hampered the development of high performance serological assays. We have now constructed an S. scabiei cDNA expression library with mRNA purified from mites isolated from red foxes. Immunoscreening of the library enabled us to clone a full-length cDNA coding for a 102.5 kDa protein. Sequence similarity searches identified the protein as a paramyosin. Recombinant S. scabiei paramyosin expressed in Escherichia coli was recognized by sera from dogs and swine infected with S. scabiei. We also designed a small paramyosin construct of about 17 kDa that included the N-terminal part, an evolutionary variable part of the helical core, and the C-terminal part of the molecule. The miniaturized protein was efficiently expressed in E. coli and was recognized by sera from immunized rabbits. These data demonstrate that the cDNA library can assist in the isolation of important S. scabiei antigens and that recombinant proteins can be useful for the study of scabies.

The Composition, Antioxidant and Antibacterial Activities of Cold-Pressed and Distilled Essential Oils of Citrus paradisi and Citrus grandis (L. Osbeck

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Ming-Chiu Ou

2015-01-01

Full Text Available The chemical composition and functional activities of cold-pressed and water distilled peel essential oils of Citrus paradisi (C. paradisi and Citrus grandis (L. Osbeck (C. grandis were investigated in present study. Yields of cold-pressed oils were much higher than those of distilled oils. Limonene was the primary ingredient of essential oils of C. paradisi (cold 92.83%; distilled 96.06% and C. grandis (cold 32.63%; distilled 55.74%. In addition, C. grandis oils obtained were rich in oxygenated or nitrogenated compounds which may be involved in reducing cardiovascular diseases or enhancing sleep effectiveness. The order of free radical scavenging activities of 4 citrus oils was distilled C. paradisi oil > cold-pressed C. paradisi oil > distilled C. grandis oil > cold-pressed C. grandis oil. Cold-pressed C. grandis oil exhibited the lowest activity in all antioxidative assays. The order of antimicrobial activities of 4 citrus oils was distilled C. grandis oil, cold-pressed C. paradisi oil > distilled C. paradisi oil > cold-pressed C. paradisi oil. Surprisingly, distilled C. grandis oil exhibited better antimicrobial activities than distilled C. paradisi oil, especially against Escherichia coli and Salmonella enterica subsp. The results also indicated that the antimicrobial activities of essential oils may not relate to their antioxidative activities.

PCR-based cDNA library construction: general cDNA libraries at the level of a few cells.

OpenAIRE

Belyavsky, A; Vinogradova, T; Rajewsky, K

1989-01-01

A procedure for the construction of general cDNA libraries is described which is based on the amplification of total cDNA in vitro. The first cDNA strand is synthesized from total RNA using an oligo(dT)-containing primer. After oligo(dG) tailing the total cDNA is amplified by PCR using two primers complementary to oligo(dA) and oligo(dG) ends of the cDNA. For insertion of the cDNA into a vector a controlled trimming of the 3' ends of the cDNA by Klenow enzyme was used. Starting from 10 J558L ...

Isolation of cDNA encoding a newly identified major allergenic protein of rye-grass pollen: intracellular targeting to the amyloplast.

Science.gov (United States)

Singh, M B; Hough, T; Theerakulpisut, P; Avjioglu, A; Davies, S; Smith, P M; Taylor, P; Simpson, R J; Ward, L D; McCluskey, J

1991-01-01

We have identified a major allergenic protein from rye-grass pollen, tentatively designated Lol pIb of 31kDa and with pI 9.0. A cDNA clone encoding Lol pIb has been isolated, sequenced, and characterized. Lol pIb is located mainly in the starch granules. This is a distinct allergen from Lol pI, which is located in the cytosol. Lol pIb is synthesized in pollen as a pre-allergen with a transit peptide targeting the allergen to amyloplasts. Epitope mapping of the fusion protein localized the IgE binding determinant in the C-terminal domain. Images PMID:1671715

Seed viability constants for Eucalyptus grandis Constantes de viabilidade para sementes de Eucalyptus grandis

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Jussara Bertho Fantinatti

2007-01-01

Full Text Available This work aimed to analyse Eucalyptus grandis W.Hill ex Maiden seed behaviour, under controlled deterioration, and to estimate viability equation constants for the species. Seeds were harvested in the growing season of 1999, and the moisture contents were adjusted from 11.3% to a range between 1.2 and 18.1% at 25ºC. The subsamples were sealed into laminate aluminium-foil packets, for storage in incubators at 40, 50 and 65±0.5ºC. The seeds presented orthodox performance, in which the constants for predicting seed longevity of E. grandis were K E = 9.661, C W = 6.467, C H = 0.03498 and C Q = 0.0002330. The usual and inverse relationship between water content and seed longevity was also observed. The lowest moisture content limit for application of the viability equation at 65ºC was 4.9%, estimated under hygroscopic equilibrium with 23% of relative humidity in the storage environment.Este trabalho teve como objetivos verificar o desempenho de sementes de Eucalyptus grandis W.Hill ex Maiden, após a deterioração em condições controladas, e obter as constantes da equação de viabilidade. As sementes foram colhidas na safra de 1999, e a umidade foi ajustada de 11,3% para valores entre 1,2 e 18,1% a 25ºC As subamostras foram acondicionadas em embalagens de alumínio termossoldadas, armazenadas a 40, 50 e 65±0,5ºC. As sementes apresentaram um desempenho ortodoxo em relação ao armazenamento. As constantes para a predição da longevidade foram K E = 9,661, C W = 6,467, C H = 0,03498 e C Q = 0,0002330. Foi observada a relação inversa entre teor de água e longevidade. O limite inferior de grau de umidade, calculado para aplicação da equação a 65ºC, foi de 4,9%, estimativa obtida sob equilíbrio higroscópico com umidade relativa de 23% no ambiente de armazenamento.

The genome of Eucalyptus grandis

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Myburg, Alexander A.; Grattapaglia, Dario; Tuskan, Gerald A.; Hellsten, Uffe; Hayes, Richard D.; Grimwood, Jane; Jenkins, Jerry; Lindquist, Erika; Tice, Hope; Bauer, Diane; Goodstein, David M.; Dubchak, Inna; Poliakov, Alexandre; Mizrachi, Eshchar; Kullan, Anand R. K.; Hussey, Steven G.; Pinard, Desre; van der Merwe, Karen; Singh, Pooja; van Jaarsveld, Ida; Silva-Junior, Orzenil B.; Togawa, Roberto C.; Pappas, Marilia R.; Faria, Danielle A.; Sansaloni, Carolina P.; Petroli, Cesar D.; Yang, Xiaohan; Ranjan, Priya; Tschaplinski, Timothy J.; Ye, Chu-Yu; Li, Ting; Sterck, Lieven; Vanneste, Kevin; Murat, Florent; Soler, Marçal; Clemente, Hélène San; Saidi, Naijib; Cassan-Wang, Hua; Dunand, Christophe; Hefer, Charles A.; Bornberg-Bauer, Erich; Kersting, Anna R.; Vining, Kelly; Amarasinghe, Vindhya; Ranik, Martin; Naithani, Sushma; Elser, Justin; Boyd, Alexander E.; Liston, Aaron; Spatafora, Joseph W.; Dharmwardhana, Palitha; Raja, Rajani; Sullivan, Christopher; Romanel, Elisson; Alves-Ferreira, Marcio; Külheim, Carsten; Foley, William; Carocha, Victor; Paiva, Jorge; Kudrna, David; Brommonschenkel, Sergio H.; Pasquali, Giancarlo; Byrne, Margaret; Rigault, Philippe; Tibbits, Josquin; Spokevicius, Antanas; Jones, Rebecca C.; Steane, Dorothy A.; Vaillancourt, René E.; Potts, Brad M.; Joubert, Fourie; Barry, Kerrie; Pappas, Georgios J.; Strauss, Steven H.; Jaiswal, Pankaj; Grima-Pettenati, Jacqueline; Salse, Jérôme; Van de Peer, Yves; Rokhsar, Daniel S.; Schmutz, Jeremy

2014-06-11

Eucalypts are the world s most widely planted hardwood trees. Their broad adaptability, rich species diversity, fast growth and superior multipurpose wood, have made them a global renewable resource of fiber and energy that mitigates human pressures on natural forests. We sequenced and assembled >94% of the 640 Mbp genome of Eucalyptus grandis into its 11 chromosomes. A set of 36,376 protein coding genes were predicted revealing that 34% occur in tandem duplications, the largest proportion found thus far in any plant genome. Eucalypts also show the highest diversity of genes for plant specialized metabolism that act as chemical defence against biotic agents and provide unique pharmaceutical oils. Resequencing of a set of inbred tree genomes revealed regions of strongly conserved heterozygosity, likely hotspots of inbreeding depression. The resequenced genome of the sister species E. globulus underscored the high inter-specific genome colinearity despite substantial genome size variation in the genus. The genome of E. grandis is the first reference for the early diverging Rosid order Myrtales and is placed here basal to the Eurosids. This resource expands knowledge on the unique biology of large woody perennials and provides a powerful tool to accelerate comparative biology, breeding and biotechnology.

Different processing of CAPA and pyrokinin precursors in the giant mealworm beetle Zophobas atratus (Tenebrionidae) and the boll weevil Anthonomus grandis grandis (Curculionidae).

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Neupert, Susanne; Marciniak, Pawel; Köhler, Rene; Nachman, Ronald J; Suh, Charles P-C; Predel, Reinhard

2018-03-01

Capa and pyrokinin (pk) genes in hexapods share a common evolutionary origin. Using transcriptomics and peptidomics, we analyzed products of these genes in two beetles, the giant mealworm beetle (Zophobas atratus; Tenebrionidae) and the boll weevil (Anthonomus grandis grandis; Curculionidae). Our data revealed that even within Coleoptera, which represents a very well-defined group of insects, highly different evolutionary developments occurred in the neuropeptidergic system. These differences, however, primarily affect the general structure of the precursors and differential processing of mature peptides and, to a lesser degree, the sequences of the active core motifs. With the differential processing of the CAPA-precursor in Z. atratus we found a perfect example of completely different products cleaved from a single neuropeptide precursor in different cells. The CAPA precursor in abdominal ganglia of this species yields primarily periviscerokinins (PVKs) whereas processing of the same precursor in neurosecretory cells of the subesophageal ganglion results in CAPA-tryptoPK and a novel CAPA-PK. Particularly important was the detection of that CAPA-PK which has never been observed in the CNS of insects before. The three different types of CAPA peptides (CAPA-tryptoPK, CAPA-PK, PVK) each represent potential ligands which activate different receptors. In contrast to the processing of the CAPA precursor from Z. atratus, no indications of a differential processing of the CAPA precursor were found in A. g. grandis. These data suggest that rapid evolutionary changes regarding the processing of CAPA precursors were still going on when the different beetle lineages diverged. The sequence of the single known PVK of A. g. grandis occupies a special position within the known PVKs of insects and might serve asa basis to develop lineage-specific peptidomimetics capable of disrupting physiological processes regulated by PVKs. Copyright © 2017 Elsevier Inc. All rights reserved.

Characterization and phylogenetic analysis of lectin gene cDNA isolated from sea cucumber ( Apostichopus japonicus) body wall

Science.gov (United States)

Xue, Zhuang; Li, Hui; Liu, Yang; Zhou, Wei; Sun, Jing; Wang, Xiuli

2017-12-01

As a `living fossil' of species origin and `rich treasure' of food and nutrition development, sea cucumber has received a lot of attentions from researchers. The cDNA library construction and EST sequencing of blood had been conducted previously in our lab. The bioinformatic analysis provided a gene fragment which is highly homologous with the genes of lectin family, named AjL ( Apostichopus japonicus lectin). To characterize and determine the phylogeny of AjL genes in early evolution, we isolated a full-length cDNA of lectin gene from the body wall of A. japonicus. The open reading frame of this gene contained 489 bp and encoded a 163 amino acids secretory protein being homologous to lectins of mammals and aquatic organisms. The deduced protein included a lectin-like domain. SDS-PAGE analysis showed that AjL migrated as a specific band (about 36.09 kDa under reducing), and agglutinated against rabbit red blood cells. AjL was similar to chain A of CEL-IV in space structure. We predicted that AjL may play the same role of CEL-IV. Our results suggested that more than one lectin gene functioned in sea cucumber and most of other species, which was fused by uncertain sequences during the evolution and encoded different proteins with diverse functions. Our findings provided the insights into the function and characteristics of lectin genes invertebrates. The results will also be helpful for the identification and structural, functional, and evolutionary analyses of lectin genes.

Cafe Grandis kõlab lautomuusika / Anneli Remme

Index Scriptorium Estoniae

Remme, Anneli, 1968-

2008-01-01

Corelli Musicu salongiõhtutel festivali "Fiesta de la Guitarra" raames esineb trio koosseisus Stewart McCoy (lautomängija), Robert Staak (lautomängija) ja Maria Staak (laulja), esinejatest. Esitatakse renessansiaja autorite lautoduette ja -laule. Kontsert 8. nov. Pärnu hotellis sarjas "Café Grandi muusikasalong"

Revisión taxonómica de Alytes grandis Brunner (Amphibia, Anura

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Sanchiz, B.

1986-12-01

Full Text Available The holotype and only existing remain referred to Alytes grandis Brunner, 1957, presumably a discoglossid frog from the German Pleistocene, is examined and compared with other living and fossil Palaearctic anurans. The analysis rejects its validity as an independent extinct species, as The material is clearly within the known variability of living Rana temporaria Linnaeus, 1758, of which it should be considered merely a sinonym.

Se examina el holotipo y único resto atribuido de Alytes grandis Brunner, 1957, un supuesto discoglósido del Pleistoceno alemán, comparándose con otros anuros actuales y fósiles del Paleártico. El análisis permite desechar su pertenencia a una especie extinta independiente, siendo en cambio atribuible con toda claridad a la viviente Rana temporaria Linnaeus, 1758, de la que debe considerarse sinónimo.
Zum systematischen Status von Alytes grandis Brunner, 1957 (Amphibia, Anura. Der bislang einzig bekannte und von Brunner (1957 beschriebene Rest zu Alytes grandis aus pleistozänen Ablagerungen der Breitenberghöhle bei Gössweinstein (Fränk, Alb wird beschrieben und aufgrund ausführlicher Vergleichsuntersuchungen der rezenten Art Rana temporaria Linnaeus, 1758, zugeordnet, Alytes grandis Brunner, 1957, ist somit als synonyrn zu Rana temporaria zu betrachten.

Isolation, nucleotide sequence and expression of a cDNA encoding feline granulocyte colony-stimulating factor.

Science.gov (United States)

Dunham, S P; Onions, D E

2001-06-21

A cDNA encoding feline granulocyte colony stimulating factor (fG-CSF) was cloned from alveolar macrophages using the reverse transcriptase-polymerase chain reaction. The cDNA is 949 bp in length and encodes a predicted mature protein of 174 amino acids. Recombinant fG-CSF was expressed as a glutathione S-transferase fusion and purified by affinity chromatography. Biological activity of the recombinant protein was demonstrated using the murine myeloblastic cell line GNFS-60, which showed an ED50 for fG-CSF of approximately 2 ng/ml. Copyright 2001 Academic Press.

JH III production, titers and degradation in relation to reproduction in male and female Anthonomus grandis.

Science.gov (United States)

Taub-Montemayor, Tina E; Min, Kyung-Jin; Chen, Zhaorigetu; Bartlett, Terri; Rankin, Mary Ann

2005-04-01

Juvenile hormone (JH) is necessary for the production of vitellogenin (Vg) in the boll weevil, Anthonomus grandis. Occurrence of Vg in this species is typically restricted to reproductively competent females, and is not detected in untreated males. However, the JH analog, methoprene stimulates Vg production in intact males and in the isolated abdomens of both male and female boll weevils (where in each case no Vg is detected without treatment), suggesting that males are competent to produce Vg but are normally not stimulated to do so. Preliminary work indicating that male boll weevil corpora allata (CA) produced little or no JH in vitro suggested that failure of males to produce Vg might be due to very low JH levels compared to females. This study re-examines the question of JH in male boll weevils by determining in vitro production of JH III by male CA during the first 10 days after adult emergence, determining hemolymph JH esterase activity during this same time period and hemolymph JH III titers in adults of both sexes. We also re-examine the ability of isolated male abdomens to produce Vg in response to hormonal stimulation, analyzing the effect of a wide range of methoprene and JH III dosages. Results indicate that male A. grandis have circulating JH titers and JH production similar to females. JH esterase activity is slightly but significantly higher in males than females. Vg production by isolated abdomens of both sexes after stimulation with methoprene or JH III was confirmed. Dose response studies indicated that high levels of methoprene were less effective than intermediate doses in stimulating Vg synthesis in both sexes. We conclude that the sexually dimorphic effect of JH on Vg synthesis is not due to differences in JH production or differences in JH titer between the sexes.

Heterogeneity of rat tropoelastin mRNA revealed by cDNA cloning

International Nuclear Information System (INIS)

Pierce, R.A.; Deak, S.B.; Stolle, C.A.; Boyd, C.D.

1990-01-01

A λgt11 library constructed from poly(A+) RNA isolated from aortic tissue of neonatal rats was screened for rat tropoelastin cDNAs. The first, screen, utilizing a human tropoelastin cDNA clone, provided rat tropoelastin cDNAs spanning 2.3 kb of carboxy-terminal coding sequence and extended into the 3'-untranslated region. A subsequent screen using a 5' rat tropoelastin cDNA clone yielded clones extending into the amino-terminal signal sequence coding region. Sequence analysis of these clones has provided the complete derived amino acid sequence of rat tropoelastin and allowed alignment and comparison with published bovine cDNA sequence. While the overall structure of rat tropoelastin is similar to bovine sequence, numerous substitutions, deletions, and insertions demonstrated considerable heterogeneity between species. In particular, the pentapeptide repeat VPGVG, characteristic of all tropoelastins analyzed to date, is replaced in rat tropoelastin by a repeating pentapeptide, IPGVG. The hexapeptide repeat VGVAPG, the bovine elastin receptor binding peptide, is not encoded by rat tropoelastin cDNAs. Variations in coding sequence between rat tropoelastin CDNA clones were also found which may represent mRNA heterogeneity produced by alternative splicing of the rat tropoelastin pre-mRNA

[Community structure of soil fauna in Eucalyptus grandis plantations at different slope locations].

Science.gov (United States)

Zhao, Yu; Zhong, Yu; Zhang, Jian; Yang, Wan-qin

2010-09-01

To understand the effects of slope location on the community structure of soil fauna in Eucalyptus grandis plantation, an investigation was made on the soil fauna in 3 E. grandis plantations at different slope locations in the hilly area of Sichuan Province from January to October 2009. A total of 39,2762 individuals were observed, belonging to 146 groups, 7 phyla, 16 classes, and 31 orders. The community composition, trophic group, diversity, and seasonal dynamics of soil fauna in the plantations all varied with slope. The abundance of macro-fauna, xeric meso- and micro-fauna, saprophagous macro-fauna, and omnivorous xeric meso- and micro-fauna increased with the decrease of slope, indicating that soil fauna had sensitive responses to the soil environmental factors affected by slope. Significant differences in the diversity of soil saprophagous macro-fauna and hygrophilous meso- and micro-fauna were observed at different slope locations, suggesting that these two faunal groups could be used as the indicators of the habitat heterogeneity of E. grandis plantations at different slope. Overall, slope location had definite effects on the community structure and distribution of soil fauna in the E. grandis plantations, but the effects were not statistically significant.

Construction and characterization of a cDNA library from human ...

African Journals Online (AJOL)

The tumor-suppressor gene p53 and its downstream genes consist of a complicated gene network, and the challenge to understand the network is to identify p53 downstream genes. In order to isolate and identify new p53 regulated genes, we constructed and characterized a normalized cDNA library from human brain ...

Transcriptome analysis in cotton boll weevil (Anthonomus grandis and RNA interference in insect pests.

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Alexandre Augusto Pereira Firmino

Full Text Available Cotton plants are subjected to the attack of several insect pests. In Brazil, the cotton boll weevil, Anthonomus grandis, is the most important cotton pest. The use of insecticidal proteins and gene silencing by interference RNA (RNAi as techniques for insect control are promising strategies, which has been applied in the last few years. For this insect, there are not much available molecular information on databases. Using 454-pyrosequencing methodology, the transcriptome of all developmental stages of the insect pest, A. grandis, was analyzed. The A. grandis transcriptome analysis resulted in more than 500.000 reads and a data set of high quality 20,841 contigs. After sequence assembly and annotation, around 10,600 contigs had at least one BLAST hit against NCBI non-redundant protein database and 65.7% was similar to Tribolium castaneum sequences. A comparison of A. grandis, Drosophila melanogaster and Bombyx mori protein families' data showed higher similarity to dipteran than to lepidopteran sequences. Several contigs of genes encoding proteins involved in RNAi mechanism were found. PAZ Domains sequences extracted from the transcriptome showed high similarity and conservation for the most important functional and structural motifs when compared to PAZ Domains from 5 species. Two SID-like contigs were phylogenetically analyzed and grouped with T. castaneum SID-like proteins. No RdRP gene was found. A contig matching chitin synthase 1 was mined from the transcriptome. dsRNA microinjection of a chitin synthase gene to A. grandis female adults resulted in normal oviposition of unviable eggs and malformed alive larvae that were unable to develop in artificial diet. This is the first study that characterizes the transcriptome of the coleopteran, A. grandis. A new and representative transcriptome database for this insect pest is now available. All data support the state of the art of RNAi mechanism in insects.

Transcriptome analysis in cotton boll weevil (Anthonomus grandis) and RNA interference in insect pests.

Science.gov (United States)

Firmino, Alexandre Augusto Pereira; Fonseca, Fernando Campos de Assis; de Macedo, Leonardo Lima Pepino; Coelho, Roberta Ramos; Antonino de Souza, José Dijair; Togawa, Roberto Coiti; Silva-Junior, Orzenil Bonfim; Pappas, Georgios Joannis; da Silva, Maria Cristina Mattar; Engler, Gilbert; Grossi-de-Sa, Maria Fatima

2013-01-01

Cotton plants are subjected to the attack of several insect pests. In Brazil, the cotton boll weevil, Anthonomus grandis, is the most important cotton pest. The use of insecticidal proteins and gene silencing by interference RNA (RNAi) as techniques for insect control are promising strategies, which has been applied in the last few years. For this insect, there are not much available molecular information on databases. Using 454-pyrosequencing methodology, the transcriptome of all developmental stages of the insect pest, A. grandis, was analyzed. The A. grandis transcriptome analysis resulted in more than 500.000 reads and a data set of high quality 20,841 contigs. After sequence assembly and annotation, around 10,600 contigs had at least one BLAST hit against NCBI non-redundant protein database and 65.7% was similar to Tribolium castaneum sequences. A comparison of A. grandis, Drosophila melanogaster and Bombyx mori protein families' data showed higher similarity to dipteran than to lepidopteran sequences. Several contigs of genes encoding proteins involved in RNAi mechanism were found. PAZ Domains sequences extracted from the transcriptome showed high similarity and conservation for the most important functional and structural motifs when compared to PAZ Domains from 5 species. Two SID-like contigs were phylogenetically analyzed and grouped with T. castaneum SID-like proteins. No RdRP gene was found. A contig matching chitin synthase 1 was mined from the transcriptome. dsRNA microinjection of a chitin synthase gene to A. grandis female adults resulted in normal oviposition of unviable eggs and malformed alive larvae that were unable to develop in artificial diet. This is the first study that characterizes the transcriptome of the coleopteran, A. grandis. A new and representative transcriptome database for this insect pest is now available. All data support the state of the art of RNAi mechanism in insects.

cDNA encoding a polypeptide including a hevein sequence

Energy Technology Data Exchange (ETDEWEB)

Raikhel, N.V.; Broekaert, W.F.; Chua, N.H.; Kush, A.

2000-07-04

A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74--79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the protein of the present invention and maltose binding protein produced by the E. coli.

cDNA encoding a polypeptide including a hevein sequence

Energy Technology Data Exchange (ETDEWEB)

Raikhel, N.V.; Broekaert, W.F.; Chua, N.H.; Kush, A.

1999-05-04

A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74--79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the protein of the present invention and maltose binding protein produced by the E. coli. 12 figs.

cDNA encoding a polypeptide including a hevein sequence

Energy Technology Data Exchange (ETDEWEB)

Raikhel, Natasha V. (Okemos, MI); Broekaert, Willem F. (Dilbeek, BE); Chua, Nam-Hai (Scarsdale, NY); Kush, Anil (New York, NY)

1999-05-04

A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the protein of the present invention and maltose binding protein produced by the E. coli.

cDNA encoding a polypeptide including a hevein sequence

Energy Technology Data Exchange (ETDEWEB)

Raikhel, N.V.; Broekaert, W.F.; Chua, N.H.; Kush, A.

1995-03-21

A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1,018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74--79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the protein of the present invention and maltose binding protein produced by the E. coli. 11 figures.

[Construction of fetal mesenchymal stem cell cDNA subtractive library].

Science.gov (United States)

Yang, Li; Wang, Dong-Mei; Li, Liang; Bai, Ci-Xian; Cao, Hua; Li, Ting-Yu; Pei, Xue-Tao

2002-04-01

To identify differentially expressed genes between fetal mesenchymal stem cell (MSC) and adult MSC, especially specified genes expressed in fetal MSC, a cDNA subtractive library of fetal MSC was constructed using suppression subtractive hybridization (SSH) technique. At first, total RNA was isolated from fetal and adult MSC. Using SMART PCR synthesis method, single-strand and double-strand cDNAs were synthesized. After Rsa I digestion, fetal MSC cDNAs were divided into two groups and ligated to adaptor 1 and adaptor 2 respectively. Results showed that the amplified library contains 890 clones. Analysis of 890 clones with PCR demonstrated that 768 clones were positive. The positive rate is 86.3%. The size of inserted fragments in these positive clones was between 0.2 - 1 kb, with an average of 400 - 600 bp. SSH is a convenient and effective method for screening differentially expressed genes. The constructed cDNA subtractive library of fetal MSC cDNA lays solid foundation for screening and cloning new and specific function related genes of fetal MSC.

Growth hormone and prolactin in Andrias davidianus: cDNA cloning, tissue distribution and phylogenetic analysis.

Science.gov (United States)

Yang, Liping; Meng, Zining; Liu, Yun; Zhang, Yong; Liu, Xiaochun; Lu, Danqi; Huang, Junhai; Lin, Haoran

2010-01-15

The Chinese giant salamander (Andrias davidianus) is one of the largest and 'living fossil' species of amphibian. To obtain genetic information for this species, the cDNAs encoding growth hormone (adGH) and prolactin (adPRL) were cloned from a pituitary cDNA library. The isolated adGH cDNA consisted of 864 bp and encoded a propeptide of 215 amino acids, while the cDNA of adPRL was 1106 bp in length and encoded a putative peptide of 229 amino acids. Expression of the GH and PRL mRNA was only detected in the pituitary. Phylogenetic analyses were performed based on the isolated pituitary hormone sequences using maximum parsimony and neighbor-joining algorithms. The clustering results are similar to that based on the morphological characteristics or the rRNA genes, which indicate that the two orders (Anura and Caudata) of amphibian were monophyletic, and that A. davidianus was diverged early in the Caudate clade. These results indicated that both the GH and PRL sequence might be useful to study the phylogenies of relatively moderate evolved groups.

Isolation and sequencing of a cDNA coding for the human DF3 breast carcinoma-associated antigen

International Nuclear Information System (INIS)

Siddiqui, J.; Abe, M.; Hayes, D.; Shani, E.; Yunis, E.; Kufe, D.

1988-01-01

The murine monoclonal antibody (mAb) DF3 reacts with a high molecular weight glycoprotein detectable in human breast carcinomas. DF3 antigen expression correlates with human breast tumor differentiation, and the detection of a cross-reactive species in human milk has suggested that this antigen might be useful as a marker of differentiated mammary epithelium. To further characterize DF3 antigen expression, the authors have isolated a cDNA clone from a λgt11 library by screening with mAb DF3. The results demonstrate that this 309-base-pair cDNA, designated pDF9.3, codes for the DF3 epitope. Southern blot analyses of EcoRI-digested DNAs from six human tumor cell lines with 32 P-labeled pDF9.3 have revealed a restriction fragment length polymorphism. Variations in size of the alleles detected by pDF9.3 were also identified in Pst I, but not in HindIII, DNA digests. Furthermore, hybridization of 32 P-labeled pDF9.3 with total cellular RNA from each of these cell lines demonstrated either one or two transcripts that varied from 4.1 to 7.1 kilobases in size. The presence of differently sized transcripts detected by pDF9.3 was also found to correspond with the polymorphic expression of DF3 glycoproteins. Nucleotide sequence analysis of pDF9.3 has revealed a highly conserved (G + C)-rich 60-base-pair tandem repeat. These findings suggest that the variation in size of alleles coding for the polymorphic DF3 glycoprotein may represent different numbers of repeats

Isolation and characterization of a cDNA clone coding for a glutathione S-transferase class delta enzyme from the biting midge Culicoides variipennis sonorensis Wirth and Jones.

Science.gov (United States)

Abdallah, M A; Pollenz, R S; Droog, F N; Nunamaker, R A; Tabachnick, W J; Murphy, K E

2000-12-01

Culicoides variipennis sonorensis is the primary vector of bluetongue viruses in North America. Glutathione S-transferases (GSTs) are enzymes that catalyze nucleophilic substitutions, converting reactive lipophilic molecules into soluble conjugates. Increased GST activity is associated with development of insecticide resistance. Described here is the isolation of the first cDNA encoding a C. variipennis GST. The clone consists of 720 translated bases encoding a protein with a M(r) of approximately 24,800 composed of 219 amino acids. The deduced amino acid sequence is similar (64%-74%) to class Delta (previously named Theta) GSTs from the dipteran genera Musca, Drosophila, Lucilia and Anopheles. The cDNA was subcloned into pET-11b, expressed in Epicurian coli BL21 (DE3) and has a specific activity of approximately 28,000 units/mg for the substrate 1-chloro-2,4-dinitrobenzene.

Normalized cDNA libraries

Science.gov (United States)

Soares, Marcelo B.; Efstratiadis, Argiris

1997-01-01

This invention provides a method to normalize a directional cDNA library constructed in a vector that allows propagation in single-stranded circle form comprising: (a) propagating the directional cDNA library in single-stranded circles; (b) generating fragments complementary to the 3' noncoding sequence of the single-stranded circles in the library to produce partial duplexes; (c) purifying the partial duplexes; (d) melting and reassociating the purified partial duplexes to moderate Cot; and (e) purifying the unassociated single-stranded circles, thereby generating a normalized cDNA library.

MODIFICACIÓN AL PROCESO DE PRODUCCIÓN MASIVA DE LOS PARASITOIDES Catolaccus grandis Y Catolaccus hunteri

Directory of Open Access Journals (Sweden)

Marco Antonio Reyes-Rosas

2010-01-01

Full Text Available El presente estudio se realizó con el objetivo de modificar la tecnología de producción masiva de Catolaccus grandis y Catolaccus hunteri, himenópteros parasitoides de Anthonomus grandis el picudo del algodonero en Tamaulipas, México en el 2002. Se determinó la eficiencia del papel toalla para el secado de manos, en el sellamiento de cápsulas moldeadas en cera de hidrocarburos y poliolefina (LCHP, donde se confina al huésped, y evaluar la efectividad del gorgojo del garbanzo Callosobruchus maculatus como hospedante facticio para la cría de ambos parasitoides. Hubo producción masiva de C. grandis y C. hunteri utilizando C. maculatus como huésped y el papel toalla fue efectivo para sellar los orificios de las cápsulas moldeadas en LCHP. La producción masiva de C. grandis y C. hunteri mediante C. maculatus, contribuyó a incrementar la seguridad y facilidad del proceso, además de una significativa reducción de costos. El papel toalla en el sellado de los orificios de las cápsulas moldeadas en LCHP, redujo en un 50% por m2 los costos del proceso, sin afectar la eficiencia del parasitismo y emergencia de C. grandis.

An alternative method for cDNA cloning from surrogate eukaryotic cells transfected with the corresponding genomic DNA.

Science.gov (United States)

Hu, Lin-Yong; Cui, Chen-Chen; Song, Yu-Jie; Wang, Xiang-Guo; Jin, Ya-Ping; Wang, Ai-Hua; Zhang, Yong

2012-07-01

cDNA is widely used in gene function elucidation and/or transgenics research but often suitable tissues or cells from which to isolate mRNA for reverse transcription are unavailable. Here, an alternative method for cDNA cloning is described and tested by cloning the cDNA of human LALBA (human alpha-lactalbumin) from genomic DNA. First, genomic DNA containing all of the coding exons was cloned from human peripheral blood and inserted into a eukaryotic expression vector. Next, by delivering the plasmids into either 293T or fibroblast cells, surrogate cells were constructed. Finally, the total RNA was extracted from the surrogate cells and cDNA was obtained by RT-PCR. The human LALBA cDNA that was obtained was compared with the corresponding mRNA published in GenBank. The comparison showed that the two sequences were identical. The novel method for cDNA cloning from surrogate eukaryotic cells described here uses well-established techniques that are feasible and simple to use. We anticipate that this alternative method will have widespread applications.

Regeneration and transformation of Eucalyptus grandis

OpenAIRE

Esteki, Leila

2013-01-01

Dissertação de mestrado em Biologia Molecular, Biotecnologia e Bioempreendedorismo em Plantas Eucalyptus grandis is the most widely used species in planted forests in tropical and subtropical areas. The traits of interest underlying Eucalyptus breeding programs concern productivity and wood quality for the pulp and paper industry, as well as biotic and abiotic stress tolerance. The development of an efficient transformation protocol is necessary to explore eucalypt resources th...

Lethal and sublethal effects of selected insecticides and an insect growth regulator on the boll weevil (Coleoptera: Curculionidae) ectoparasitoid Catolaccus grandis (Hymenoptera: Pteromalidae).

Science.gov (United States)

Elzen, G W; Maldonado, S N; Rojas, M G

2000-04-01

A laboratory culture of Catolaccus grandis (Burks), an ectoparasitoid of the boll weevil, Anthonomus grandis grandis Boheman, was exposed to lethal and sublethal doses of insecticides and an insect growth regulator using a spray chamber bioassay. Materials tested were azinphos-methyl, endosulfan, fipronil, malathion, cyfluthrin, dimethoate, spinosad, methyl parathion, acephate, oxamyl, and tebufenozide. At full rates, spinosad was significantly less toxic to female C. grandis than other treatments except endosulfan. Fipronil and malathion were significantly more toxic to females than other treatments. Most of the chemicals tested were highly toxic to male C. grandis; spinosad was least toxic. At reduced rates, most of 4 selected chemicals tested were low in toxicity to C. grandis; however, a reduced rate of malathion was significantly more toxic to females than other treatments. No C. grandis pupae developed from parasitism during a 24-h treatment period with malathion or spinosad. The sex ratio of progeny from sprayed adults appeared to be unaffected by the treatments.

Two human cDNA molecules coding for the Duchenne muscular dystrophy (DMD) locus are highly homologous

Energy Technology Data Exchange (ETDEWEB)

Rosenthal, A.; Speer, A.; Billwitz, H. (Zentralinstitut fuer Molekularbiologie, Berlin-Buch (Germany Democratic Republic)); Cross, G.S.; Forrest, S.M.; Davies, K.E. (Univ. of Oxford (England))

1989-07-11

Recently the complete sequence of the human fetal cDNA coding for the Duchenne muscular dystrophy (DMD) locus was reported and a 3,685 amino acid long, rod-shaped cytoskeletal protein (dystrophin) was predicted as the protein product. Independently, the authors have isolated and sequenced different DMD cDNA molecules from human adult and fetal muscle. The complete 12.5 kb long sequence of all their cDNA clones has now been determined and they report here the nucleotide (nt) and amino acid (aa) differences between the sequences of both groups. The cDNA sequence comprises the whole coding region but lacks the first 110 nt from the 5{prime}-untranslated region and the last 1,417 nt of the 3{prime}-untranslated region. They have found 11 nt differences (approximately 99.9% homology) from which 7 occurred at the aa level.

cDNA cloning, mRNA distribution and heterogeneity, chromosomal location, and RFLP analysis of human osteopontin (OPN)

DEFF Research Database (Denmark)

Young, M F; Kerr, J M; Termine, J D

1990-01-01

A human osteopontin (OP) cDNA was isolated from a library made from primary cultures of human bone cells. The distribution of osteopontin mRNA in human tissues was investigated by Northern analysis and showed that the human message was predominant in cultures of bone cells and in decidua cells...... osteopontin cDNA indicated that the gene is a single copy with an approximate length of 5.4-8.2 kb....

Anthraquinones and naphtoquinones from the bark of Tectona grandis (Verbenaceae) reforestation specimen

OpenAIRE

Moreira, Rafael Y.O.; Arruda, Mara S.P.; Arruda, Alberto C.; Santos, Lourivaldo S.; Müller, Adolfo H.; Guilhon, Giselle M.S.P.; Santos, Alberdan S.; Terezo, Evaristo

2006-01-01

O fracionamento do extrato hexânico do caule de um espécime de reflorestamento de Tectona grandis (Verbenaceae), através de procedimentos fitoquímicos clássicos, levou ao isolamento das naftoquinonas lapachol e desidro-a-lapachona e das antraquinonas tectoquinona e obtusifolina. As estruturas das substâncias foram caracterizadas através da análise de métodos espectrométricos de RMN. Este é o primeiro estudo fitoquímico de um espécime de reflorestamento de Tectona grandis, no Brasil, sendo o o...

Stomatal characteristics of Eucalyptus grandis clonal hybrids in ...

African Journals Online (AJOL)

This study describes the stomatal response occurring during water stress and subsequent recovery of three Eucalyptus grandis clonal hybrids. The aim was to investigate the degree to which stomatal conductance (gs) and stomatal density differ between the clonal hybrids across seasons and in response to water stress.

Morphology, diet, and temperature dependent host-free survival of the boll weevil, Anthonomus grandis (Coleoptera: Curculionidae)

Science.gov (United States)

The boll weevil, Anthonomus grandis grandis Boheman, is an important pest of cotton (Gossypium spp.) in South America, Mexico, and southernmost Texas in the United States. A key factor in the persistence of the boll weevil is its ability to survive the non-cotton season. Mechanisms facilitating this...

Contenido de los nutrientes básicos en Catolaccus grandis Burks criados sobre larvas del picudo del algodon Basic nutrients content of Catolaccus grandis Burks reared in cotton boll weevil larvae

Directory of Open Access Journals (Sweden)

LÚCIA HELENA AVELINO ARAUJO

2000-09-01

Full Text Available El objetivo del trabajo fue determinar los niveles de carbohidratos, proteínas solubles y aminoácidos libres de larvas, pupas hembras y adultos hembras de Catolaccus grandis (Burks (Hymenoptera: Pteromalidae criados sobre larvas del picudo del algodón envenenadas por hembras del parasitoide y por larvas de primer instar del parasitoide. Esto estudio fue conducido en la Unidade de Investigación de Control Biologico de Plagas del Departamento de Agricultura de los Estados Unidos de la América, en Weslaco, Texas. Las 20 muestras de cada uno de los tres estados de desarrollo: tercer instar larval, pupas hembra y adultos hembra del parasitoide C. grandis, fueron separadas y pesadas individualmente y se cuantió el contenido de carbohidratos totales, proteínas solubles totales y aminoácidos libres criados en diferentes sustratos. Los resultados obtenidos confirman la existencia de patrones metabólicos significativamente distintos de estos nutrientes básicos.The aim of this work was to determine the levels of carbohydrate, soluble proteins and free amino acids of larvae, pupae and adult females Catolaccus grandis Burks (Hymenoptera: Pteromalidae which were reared in cotton (Gossypium hirsutum L. r. latifolium Hutch boll weevil (Anthonomus grandis larvae venomized by ectoparasitoid of female and 1st instar ectoparasitoid larvae. This study was carried out at the Biological Control of Pests Research Unit, Weslaco, Texas. The twenty samples of each one of three stages of development: 3rd instar larval, female pupae and female adult of parasitoid C. grandis were separated and individually weighted, and levels of carbohydrates, proteins and amino acids were quantified when reared in different substrates. The results confirmed the existence of metabolic patterns significantly distinct from the basic nutrient model.

Intercropping Acacia mangium stimulates AMF colonization and soil phosphatase activity in Eucalyptus grandis

Directory of Open Access Journals (Sweden)

Daniel Bini

Full Text Available ABSTRACT: Arbuscular mycorrhizal fungi (AMF are very important to plant nutrition, mostly in terms of acquisition of P and micronutrients. While Acacia mangium is closely associated with AMF throughout the whole cycle, Eucalyptus grandis presents this symbiosis primarily at the seedling stage. The aim of this study was to evaluate the dynamics of AMF in these two tree species in both pure and mixed plantations during the first 20 months after planting. We evaluated the abundance, richness and diversity of AMF spores, the rate of AMF mycorrhizal root colonization, enzymatic activity and soil and litter C, N and P. There was an increase in AMF root colonization of E. grandis when intercropped with A. mangium as well as an increase in the activity of acid and alkaline phosphatase in the presence of leguminous trees. AMF colonization and phosphatase activities were both involved in improvements in P cycling and P nutrition in soil. In addition, P cycling was favored in the intercropped plantation, which showed negative correlation with litter C/N and C/P ratios and positive correlation with soil acid phosphatase activity and soil N and P concentrations. Intercropping A. mangium and E. grandis maximized AMF root colonization of E. grandis and phosphatase activity in the soil, both of which accelerate P cycling and forest performance.

Genetic improvement of Eucalyptus grandis using breeding seedling ...

African Journals Online (AJOL)

Eucalyptus grandis is commercially important in Zimbabwe and a breeding program has been in progress since 1962. A classical breeding strategy was used initially but, in 1981, the Multiple Population Breeding Strategy (MPBS) was implemented and the concept of the Breeding Seedling Orchard (BSO) became central to ...

Primeiro registro de Oligonychus yothersi (McGregor (Acari: Tetranychidae em Eucalyptus grandis Hill ex Maiden no Brasil First record of Oligonychus yothersi (McGregor (Acari: Tetranychidae on Eucalyptus grandis Hill ex Maiden in Brazil

Directory of Open Access Journals (Sweden)

Fabrício Fagundes Pereira

2005-08-01

Full Text Available Relata-se a infestação de um ácaro-vermelho em mudas clonais de Eucalyptus grandis Hill ex Maiden, mantidas em casa de vegetação no município de Martinho Campos, Minas Gerais. O ácaro foi observado na parte superior das folhas que exibiam sinais de sucção de seiva e bronzeamento. Essas injúrias causaram desenvolvimento anormal e morte de plantas. O ácaro foi identificado como Oligonychus yothersi (McGregor (Acari: Tetranychidae, e isso representa o primeiro registro dessa espécie em mudas clonais de E. grandis no Brasil.An infestation of the red spider mite was reported in clone seedlings of Eucalyptus grandis Hill ex Maiden under greenhouse conditions, in the municipality of Martinho Campos, Minas Gerais State. The spider mite was found on the leaf upper faces with signs of sap suction and bronzing. Such injuries caused abnormal development and plant death. The spider mite was identified as Oligonychus yothersi (McGregor (Acari: Tetranychidae. This is the first record of O. yothersi on E. grandis seedlings in Brazil.

cDNA sequence quality data - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available List Contact us Budding yeast cDNA sequencing project cDNA sequence quality data Data detail Data name cDNA sequence quality... data DOI 10.18908/lsdba.nbdc00838-003 Description of data contents Phred's quality score. P...tion Download License Update History of This Database Site Policy | Contact Us cDNA sequence quality

cDNA encoding a polypeptide including a hev ein sequence

Energy Technology Data Exchange (ETDEWEB)

Raikhel, Natasha V. (Okemos, MI); Broekaert, Willem F. (Dilbeek, BE); Chua, Nam-Hai (Scarsdale, NY); Kush, Anil (New York, NY)

2000-07-04

A cDNA clone (HEV1) encoding hevein was isolated via polymerase chain reaction (PCR) using mixed oligonucleotides corresponding to two regions of hevein as primers and a Hevea brasiliensis latex cDNA library as a template. HEV1 is 1018 nucleotides long and includes an open reading frame of 204 amino acids. The deduced amino acid sequence contains a putative signal sequence of 17 amino acid residues followed by a 187 amino acid polypeptide. The amino-terminal region (43 amino acids) is identical to hevein and shows homology to several chitin-binding proteins and to the amino-termini of wound-induced genes in potato and poplar. The carboxyl-terminal portion of the polypeptide (144 amino acids) is 74-79% homologous to the carboxyl-terminal region of wound-inducible genes of potato. Wounding, as well as application of the plant hormones abscisic acid and ethylene, resulted in accumulation of hevein transcripts in leaves, stems and latex, but not in roots, as shown by using the cDNA as a probe. A fusion protein was produced in E. coli from the protein of the present invention and maltose binding protein produced by the E. coli.

Cloning and cDNA sequence of the dihydrolipoamide dehydrogenase component of human α-ketoacid dehydrogenase complexes

International Nuclear Information System (INIS)

Pons, G.; Raefsky-Estrin, C.; Carothers, D.J.; Pepin, R.A.; Javed, A.A.; Jesse, B.W.; Ganapathi, M.K.; Samols, D.; Patel, M.S.

1988-01-01

cDNA clones comprising the entire coding region for human dihydrolipoamide dehydrogenase have been isolated from a human liver cDNA library. The cDNA sequence of the largest clone consisted of 2082 base pairs and contained a 1527-base open reading frame that encodes a precursor dihydrolipoamide dehydrogenase of 509 amino acid residues. The first 35-amino acid residues of the open reading frame probably correspond to a typical mitochondrial import leader sequence. The predicted amino acid sequence of the mature protein, starting at the residue number 36 of the open reading frame, is almost identical (>98% homology) with the known partial amino acid sequence of the pig heart dihydrolipoamide dehydrogenase. The cDNA clone also contains a 3' untranslated region of 505 bases with an unusual polyadenylylation signal (TATAAA) and a short poly(A) track. By blot-hybridization analysis with the cDNA as probe, two mRNAs, 2.2 and 2.4 kilobases in size, have been detected in human tissues and fibroblasts, whereas only one mRNA (2.4 kilobases) was detected in rat tissues

EFFECT OF FERTILIZATION ON MECHANICAL PROPERTIES OF THE WOOD OF Eucalyptus grandis

Directory of Open Access Journals (Sweden)

Israel Luiz de Lima

2011-09-01

Full Text Available The effect of the fertilization in the amount and quality of the produced wood is one of the questions to be considered in the research of the Eucalyptus grandis. The present work aimed to evaluate the fertilization effect in the mechanical properties of Eucalyptus grandis. The population of Eucalyptus grandis was 21 years old and was managed under the system of selective thinning, with application of fertilizers. The factors used in this study were: presence or absence of fertilizers, two positions of log and five radial positions. The influences of the factors and of their combinations were evaluated regarding to compression strength, shear strength, modulus of rupture and modulus of elasticity in static banding. The compressive strength and the modulus of elasticity had been influenced by the factors: fertilizer and radial positions of the log. There was also an increase in the direction of the pith-bark in all studied properties. A good positive relationship was found to exist among the compression strength, the shear, the modulus of rupture and the modulus of elasticity with radial position.

Diagnosing foliar nutrient dynamics of Eucalyptus grandis in ...

African Journals Online (AJOL)

Fertilisation is one of the most cost-effective methods of increasing and maintaining the productivity of Eucalyptus grandis plantations in South Africa. This silvicultural practice can be optimised by using the foliar nutrient ratios measured in plants at maximum growth as a guideline for fertiliser application. The foliar nutrient ...

Isolation and characterisation of cDNA clones representing the genes encoding the major tuber storage protein (dioscorin) of yam (Dioscorea cayenensis Lam.).

Science.gov (United States)

Conlan, R S; Griffiths, L A; Napier, J A; Shewry, P R; Mantell, S; Ainsworth, C

1995-06-01

cDNA clones encoding dioscorins, the major tuber storage proteins (M(r) 32,000) of yam (Dioscorea cayenesis) have been isolated. Two classes of clone (A and B, based on hybrid release translation product sizes and nucleotide sequence differences) which are 84.1% similar in their protein coding regions, were identified. The protein encoded by the open reading frame of the class A cDNA insert is of M(r) 30,015. The difference in observed and calculated molecular mass might be attributed to glycosylation. Nucleotide sequencing and in vitro transcription/translation suggest that the class A dioscorin proteins are synthesised with signal peptides of 18 amino acid residues which are cleaved from the mature peptide. The class A and class B proteins are 69.6% similar with respect to each other, but show no sequence identity with other plant proteins or with the major tuber storage proteins of potato (patatin) or sweet potato (sporamin). Storage protein gene expression was restricted to developing tubers and was not induced by growth conditions known to induce expression of tuber storage protein genes in other plant species. The codon usage of the dioscorin genes suggests that the Dioscoreaceae are more closely related to dicotyledonous than to monocotyledonous plants.

Modelos de predição para sobrevivência de plantas de Eucalyptus grandis Prediction models of Eucalyptus grandis plant survival

Directory of Open Access Journals (Sweden)

Telde Natel Custódio

2009-01-01

Full Text Available Objetivou-se com este trabalho comparar modelos de predição de plantas sobreviventes de Eucalyptus grandis. Utilizaram-se os seguintes modelos: modelo linear misto com os dados transformados, utilizando-se as transformações angular e BOX-COX; modelo linear generalizado misto com distribuição binomial e funções de ligação logística, probit e complemento log-log; modelo linear generalizado misto com distribuição Poisson e função de ligação logarítmica. Os dados são provenientes de um experimento em blocos ao acaso, para avaliação de progênies maternas de Eucalyptus grandis, aos 5 anos de idade, em que a variável resposta são plantas sobreviventes. Para comparação dos efeitos entre os modelos foram estimadas as correlações de Spearman e aplicado o teste de permutação de Fisher. Foi possível concluir que, o modelo linear generalizado misto com distribuição Poisson e função de ligação logarítmica se ajustou mal aos dados e que as estimativas para os efeitos fixos e predição para os efeitos aleatórios, não se diferenciaram entre os demais modelos estudados.The objective of this work was to compare models for prediction of the survival of plants of Eucalyptus grandis. The following models were used: linear mixed model with the transformed data, by utilizing the angular transformations and BOX-COX; generalized linear mixed model with binomial distribution and logistic functions, probit and complement log-log links; generalized linear mixed model with Poisson distribution and logarithmic link function. The data came from a randomized block experiment for evaluation of Eucalyptus grandis maternal progenies at five years old, in which the variable response are surviving plants. For comparison of the effects among the models the correlations of Spearman were estimated and the test of permutation of Fisher was applied. It was possible to conclude that: the generalized linear mixed model with Poisson distribution and

Isolation and characterization of cDNA encoding the 80-kDa subunit protein of the human autoantigen Ku (p70/p80) recognized by autoantibodies from patients with scleroderma-polymyositis overlap syndrome

International Nuclear Information System (INIS)

Mimori, Tsuneyo; Ohosone, Yasuo; Hama, Nobuaki; Suwa, Akira; Akizuki, Masashi; Homma, Mitsuo; Griffith, A.J.; Hardin, J.A.

1990-01-01

Anti-Ku (p70/p80) autoantibodies in patients with scleroderma-polymyositis overlap syndrome recognize a 70-kDa/80-kDa protein heterodimer which binds to terminal regions of double-stranded DNA. In the present study, the authors isolated full-length cDNAs that encode the 80-kDa Ku subunit. Initial screening of a human spleen cDNA library with anti-Ku antibodies yielded a cDNA of 1.0 kilobase (kb) (termed K71) encoding a portion of the 80-kDa Ku polypeptide (identification based on immunological criteria). In RNA blots, this cDNA hybridized with two mRNAs of 3.4 and 2.6 kb. In vitro transcription and translation experiments produced an immunoprecipitable polypeptide which comigrated with the 80-kDa Ku subunit. The Ku80-6 cDNA proved to be 3304 nucleotides in length, with an additional poly(A) tail, closely approximating the size of the larger mRNA. It contains a single long open reading frame encoding 732 amino acids. The putative polypeptide has a high content of acidic amino acids and a region with periodic repeat of leucine in every seventh position which may form the leucine zipper structure. In genomic DNA blots, probes derived from the opposite ends of cDNA Ku80-6 hybridized with several nonoverlapping restriction fragments from human leukocyte DNA, indicating that the gene encoding the 80-kDa Ku polypeptide is divided into several exons by intervening sequences

cDNA sequence of human transforming gene hst and identification of the coding sequence required for transforming activity

International Nuclear Information System (INIS)

Taira, M.; Yoshida, T.; Miyagawa, K.; Sakamoto, H.; Terada, M.; Sugimura, T.

1987-01-01

The hst gene was originally identified as a transforming gene in DNAs from human stomach cancers and from a noncancerous portion of stomach mucosa by DNA-mediated transfection assay using NIH3T3 cells. cDNA clones of hst were isolated from the cDNA library constructed from poly(A) + RNA of a secondary transformant induced by the DNA from a stomach cancer. The sequence analysis of the hst cDNA revealed the presence of two open reading frames. When this cDNA was inserted into an expression vector containing the simian virus 40 promoter, it efficiently induced the transformation of NIH3T3 cells upon transfection. It was found that one of the reading frames, which coded for 206 amino acids, was responsible for the transforming activity

Cloning, sequencing and expression of a novel xylanase cDNA from ...

African Journals Online (AJOL)

STORAGESEVER

2008-12-03

Dec 3, 2008 ... First strand cDNA was synthesized by RT-PCR with Oligo(dT)15 using mRNA isolated ... 4°C. Single colonies were picked into 5 mL BMGY medium for preculture, and incubated ... to fold properly into a native conformation. Without the .... polymorphism is often used in taxonomy, but now, it is being well ...

Isolation and characterisation of the cDNA encoding a glycosylated accessory protein of pea chloroplast DNA polymerase.

OpenAIRE

Gaikwad, A; Tewari, K K; Kumar, D; Chen, W; Mukherjee, S K

1999-01-01

The cDNA encoding p43, a DNA binding protein from pea chloroplasts (ct) that binds to cognate DNA polymerase and stimulates the polymerase activity, has been cloned and characterised. The characteristic sequence motifs of hydroxyproline-rich glyco-proteins (HRGP) are present in the cDNA corres-ponding to the N-terminal domain of the mature p43. The protein was found to be highly O-arabinosylated. Chemically deglycosylated p43 (i.e. p29) retains its binding to both DNA and pea ct-DNA polymeras...

[Primary culture of cat intestinal epithelial cell and construction of its cDNA library].

Science.gov (United States)

Ye, L; Gui-Hua, Z; Kun, Y; Hong-Fa, W; Ting, X; Gong-Zhen, L; Wei-Xia, Z; Yong, C

2017-04-12

Objective To establish the primary cat intestinal epithelial cells (IECs) culture methods and construct the cDNA library for the following yeast two-hybrid experiment, so as to screen the virulence interaction factors among the final host. Methods The primary cat IECs were cultured by the tissue cultivation and combined digestion with collagenase XI and dispase I separately. Then the cat IECs cultured was identified with the morphological observation and cyto-keratin detection, by using goat anti-cyto-keratin monoclonal antibodies. The mRNA of cat IECs was isolated and used as the template to synthesize the first strand cDNA by SMART™ technology, and then the double-strand cDNAs were acquired by LD-PCR, which were subsequently cloned into the plasmid PGADT7-Rec to construct yeast two-hybrid cDNA library in the yeast strain Y187 by homologous recombination. Matchmaker™ Insert Check PCR was used to detect the size distribution of cDNA fragments after the capacity calculation of the cDNA library. Results The comparison of the two cultivation methods indicated that the combined digestion of collagenase XI and dispase I was more effective than the tissue cultivation. The cat IECs system of continuous culture was established and the cat IECs with high purity were harvested for constructing the yeast two-hybrid cDNA library. The library contained 1.1×10 6 independent clones. The titer was 2.8×10 9 cfu/ml. The size of inserted fragments was among 0.5-2.0 kb. Conclusion The yeast two-hybrid cDNA library of cat IECs meets the requirements of further screen research, and this study lays the foundation of screening the Toxoplasma gondii virulence interaction factors among the cDNA libraries of its final hosts.

cDNA sequence analysis of a 29-kDa cysteine-rich surface antigen of pathogenic Entamoeba histolytica

International Nuclear Information System (INIS)

Torian, B.E.; Stroeher, V.L.; Stamm, W.E.; Flores, B.M.; Hagen, F.S.

1990-01-01

A λgt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. A 29-kDa protein was identified as a surface antigen when mAbs were used to immunoprecipitate the antigen from metabolically 35 S-labeled live trophozoites. The surface location of the antigen was corroborated by mAb immunoprecipitation of a 29-kDa protein from surface- 125 I-labeled whole trophozoites as well as by the reaction of mAbs with live trophozoites in an indirect immunofluorescence assay performed at 4 degree C. Immunoblotting with mAbs demonstrated that the antigen was present on four axenic isolates tested. mAbs recognized epitopes on the 29-kDa native antigen on some but not all clinical isolates tested

cDNA sequence analysis of a 29-kDa cysteine-rich surface antigen of pathogenic Entamoeba histolytica

Energy Technology Data Exchange (ETDEWEB)

Torian, B.E.; Stroeher, V.L.; Stamm, W.E. (Univ. of Washington, Seattle (USA)); Flores, B.M. (Louisiana State Univ. Medical Center, New Orleans (USA)); Hagen, F.S. (Zymogenetics Incorporated, Seattle, WA (USA))

1990-08-01

A {lambda}gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNA probe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein was used to generate 22 monoclonal antibodies (mAbs) and a mouse polyclonal antiserum specific for the E. histolytica portion of the fusion protein. A 29-kDa protein was identified as a surface antigen when mAbs were used to immunoprecipitate the antigen from metabolically {sup 35}S-labeled live trophozoites. The surface location of the antigen was corroborated by mAb immunoprecipitation of a 29-kDa protein from surface-{sup 125}I-labeled whole trophozoites as well as by the reaction of mAbs with live trophozoites in an indirect immunofluorescence assay performed at 4{degree}C. Immunoblotting with mAbs demonstrated that the antigen was present on four axenic isolates tested. mAbs recognized epitopes on the 29-kDa native antigen on some but not all clinical isolates tested.

Display of a Maize cDNA library on baculovirus infected insect cells

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Jones Ian M

2008-08-01

Full Text Available Abstract Background Maize is a good model system for cereal crop genetics and development because of its rich genetic heritage and well-characterized morphology. The sequencing of its genome is well advanced, and new technologies for efficient proteomic analysis are needed. Baculovirus expression systems have been used for the last twenty years to express in insect cells a wide variety of eukaryotic proteins that require complex folding or extensive posttranslational modification. More recently, baculovirus display technologies based on the expression of foreign sequences on the surface of Autographa californica (AcMNPV have been developed. We investigated the potential of a display methodology for a cDNA library of maize young seedlings. Results We constructed a full-length cDNA library of young maize etiolated seedlings in the transfer vector pAcTMVSVG. The library contained a total of 2.5 × 105 independent clones. Expression of two known maize proteins, calreticulin and auxin binding protein (ABP1, was shown by western blot analysis of protein extracts from insect cells infected with the cDNA library. Display of the two proteins in infected insect cells was shown by selective biopanning using magnetic cell sorting and demonstrated proof of concept that the baculovirus maize cDNA display library could be used to identify and isolate proteins. Conclusion The maize cDNA library constructed in this study relies on the novel technology of baculovirus display and is unique in currently published cDNA libraries. Produced to demonstrate proof of principle, it opens the way for the development of a eukaryotic in vivo display tool which would be ideally suited for rapid screening of the maize proteome for binding partners, such as proteins involved in hormone regulation or defence.

Characterization of cDNA for human tripeptidyl peptidase II: The N-terminal part of the enzyme is similar to subtilisin

International Nuclear Information System (INIS)

Tomkinson, B.; Jonsson, A-K

1991-01-01

Tripeptidyl peptidase II is a high molecular weight serine exopeptidase, which has been purified from rat liver and human erythrocytes. Four clones, representing 4453 bp, or 90% of the mRNA of the human enzyme, have been isolated from two different cDNA libraries. One clone, designated A2, was obtained after screening a human B-lymphocyte cDNA library with a degenerated oligonucleotide mixture. The B-lymphocyte cDNA library, obtained from human fibroblasts, were rescreened with a 147 bp fragment from the 5' part of the A2 clone, whereby three different overlapping cDNA clones could be isolated. The deduced amino acid sequence, 1196 amino acid residues, corresponding to the longest open rading frame of the assembled nucleotide sequence, was compared to sequences of current databases. This revealed a 56% similarity between the bacterial enzyme subtilisin and the N-terminal part of tripeptidyl peptidase II. The enzyme was found to be represented by two different mRNAs of 4.2 and 5.0 kilobases, respectively, which probably result from the utilziation of two different polyadenylation sites. Futhermore, cDNA corresponding to both the N-terminal and C-terminal part of tripeptidyl peptidase II hybridized with genomic DNA from mouse, horse, calf, and hen, even under fairly high stringency conditions, indicating that tripeptidyl peptidase II is highly conserved

Shoot and root morphogenesis from Eucalyptus grandis x urophylla ...

African Journals Online (AJOL)

Eucalyptus grandis x urophylla plantlets were regenerated via indirect organogenesis. Histological assessment of their development focused on identifying the calli, the differentiation of shoots from the calli and the shoot-root junction from the nascent shoots. Vascular tissue formation within the callus preceded that of ...

A Newly Identified Passive Hyperaccumulator Eucalyptus grandis × E. urophylla under Manganese Stress.

Science.gov (United States)

Xie, Qingqing; Li, Zhenji; Yang, Limin; Lv, Jing; Jobe, Timothy O; Wang, Qiuquan

2015-01-01

Manganese (Mn) is an essential micronutrient needed for plant growth and development, but can be toxic to plants in excess amounts. However, some plant species have detoxification mechanisms that allow them to accumulate Mn to levels that are normally toxic, a phenomenon known as hyperaccumulation. These species are excellent candidates for developing a cost-effective remediation strategy for Mn-polluted soils. In this study, we identified a new passive Mn-hyperaccumulator Eucalyptus grandis × E. urophylla during a field survey in southern China in July 2010. This hybrid can accumulate as much as 13,549 mg/kg DW Mn in its leaves. Our results from Scanning Electron Microscope (SEM) X-ray microanalysis indicate that Mn is distributed in the entire leaf and stem cross-section, especially in photosynthetic palisade, spongy mesophyll tissue, and stem xylem vessels. Results from size-exclusion chromatography coupled with ICP-MS (Inductively coupled plasma mass spectrometry) lead us to speculate that Mn associates with relatively high molecular weight proteins and low molecular weight organic acids, including tartaric acid, to avoid Mn toxicity. Our results provide experimental evidence that both proteins and organic acids play important roles in Mn detoxification in Eucalyptus grandis × E. urophylla. The key characteristics of Eucalyptus grandis × E. urophylla are an increased Mn translocation facilitated by transpiration through the xylem to the leaves and further distribution throughout the leaf tissues. Moreover, the Mn-speciation profile obtained for the first time in different cellular organelles of Eucalyptus grandis × E. urophylla suggested that different organelles have differential accumulating abilities and unique mechanisms for Mn-detoxification.

Soil carbon estimation from eucalyptus grandis using canopy spectra

African Journals Online (AJOL)

Mapping soil fertility parameters, such as soil carbon (C), is fundamentally important for forest management and research related to forest growth and climate change. This study seeks to establish the link between Eucalyptus grandis canopy spectra and soil carbon using raw and continuum-removed spectra. Canopy-level ...

Molecular cloning of a human glycophorin B cDNA: nucleotide sequence and genomic relationship to glycophorin A

International Nuclear Information System (INIS)

Siebert, P.D.; Fukuda, M.

1987-01-01

The authors describe the isolation and nucleotide sequence of a human glycophorin B cDNA. The cDNA was identified by differential hybridization of synthetic oligonucleotide probes to a human erythroleukemic cell line (K562) cDNA library constructed in phage vector λgt10. The nucleotide sequence of the glycophorin B cDNA was compared with that of a previously cloned glycophorin A cDNA. The nucleotide sequences encoding the NH 2 -terminal leader peptide and first 26 amino acids of the two proteins are nearly identical. This homologous region is followed by areas specific to either glycophorin A or B and a number of small regions of homology, which in turn are followed by a very homologous region encoding the presumed membrane-spanning portion of the proteins. They used RNA blot hybridization with both cDNA and synthetic oligonucleotide probes to prove our previous hypothesis that glycophorin B is encoded by a single 0.5- to 0.6-kb mRNA and to show that glycophorins A and B are negatively and coordinately regulated by a tumor-promoting phorbol ester, phorbol 12-myristate 13-acetate. They established the intron/exon structure of the glycophorin A and B genes by oligonucleotide mapping; the results suggest a complex evolution of the glycophorin genes

Population structure and genetic diversity of the boll weevil, Anthonomus grandis (Coleoptera: Curculionidae), on Gossypium in North America

Science.gov (United States)

While the boll weevil, Anthonomus grandis, has been identified as one of the most devastating pests in U.S. history, its origin and activity in Mexico, both on wild and cultivated cotton hosts (genus Gossypium), is poorly understood. Three forms (geographical or host-associated races) of A. grandis ...

Water relations of Eucalyptus nitens x Eucalyptus grandis : is there ...

African Journals Online (AJOL)

Water relations of Eucalyptus nitens x Eucalyptus grandis : is there interclonal variation in response to experimentally imposed water stress? ... Southern Forests: a Journal of Forest Science ... However, water stress reduced shoot hydraulic conductance and stem hydraulic conductivity with significant interclonal effects.

Construction and characterization of the alpha form of a cardiac myosin heavy chain cDNA clone and its developmental expression in the Syrian hamster.

OpenAIRE

Liew, C C; Jandreski, M A

1986-01-01

A cDNA clone, pVHC1, was isolated from a Syrian hamster heart cDNA library and was compared to the rat alpha (pCMHC21) and beta (pCMHC5) ventricular myosin heavy chain cDNA clones. The DNA sequence and amino acid sequence deducted from the DNA show more homology with pCMHC21 than pCMHC5. This indicates that pVHC1 is an alpha ventricular myosin heavy chain cDNA clone. However, even though pVHC1 shows a high degree of nucleotide and amino acid conservation with the rat myosin heavy chain sequen...

The cDNA sequence of a neutral horseradish peroxidase.

Science.gov (United States)

Bartonek-Roxå, E; Eriksson, H; Mattiasson, B

1991-02-16

A cDNA clone encoding a horseradish (Armoracia rusticana) peroxidase has been isolated and characterized. The cDNA contains 1378 nucleotides excluding the poly(A) tail and the deduced protein contains 327 amino acids which includes a 28 amino acid leader sequence. The predicted amino acid sequence is nine amino acids shorter than the major isoenzyme belonging to the horseradish peroxidase C group (HRP-C) and the sequence shows 53.7% identity with this isoenzyme. The described clone encodes nine cysteines of which eight correspond well with the cysteines found in HRP-C. Five potential N-glycosylation sites with the general sequence Asn-X-Thr/Ser are present in the deduced sequence. Compared to the earlier described HRP-C this is three glycosylation sites less. The shorter sequence and fewer N-glycosylation sites give the native isoenzyme a molecular weight of several thousands less than the horseradish peroxidase C isoenzymes. Comparison with the net charge value of HRP-C indicates that the described cDNA clone encodes a peroxidase which has either the same or a slightly less basic pI value, depending on whether the encoded protein is N-terminally blocked or not. This excludes the possibility that HRP-n could belong to either the HRP-A, -D or -E groups. The low sequence identity (53.7%) with HRP-C indicates that the described clone does not belong to the HRP-C isoenzyme group and comparison of the total amino acid composition with the HRP-B group does not place the described clone within this isoenzyme group. Our conclusion is that the described cDNA clone encodes a neutral horseradish peroxidase which belongs to a new, not earlier described, horseradish peroxidase group.

Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: Homology to Epstein-Barr virus open reading frame BCRFI

Energy Technology Data Exchange (ETDEWEB)

Vieira, P.; De Waal-Malefyt, R.; Dang, M.N.; Johnson, K.E.; Kastelein, R.; Fiorentino, D.F.; DeVries, J.E.; Roncarolo, M.G.; Mosmann, T.R.; Moore, K.W. (DNAX Research Inst. of Molecular and Cellular Biology, Palo Alto, CA (United States))

1991-02-15

The authors demonstrated the existence of human cytokine synthesis inhibitory factor (DSIF) (interleukin 10 (IL-10)). cDNA clones encoding human IL-10 (hIL-10) were isolated from a tetanus toxin-specific human T-cell clone. Like mouse IL-10, hIL-10 exhibits strong DNA and amino acid sequence homology to an open reading frame in the Epstein-Barr virus, BDRFL. hIL-10 and the BCRFI product inhibit cytokine synthesis by activated human peripheral blood mononuclear cells and by a mouse Th1 clone. Both hIL-10 and mouse IL-10 sustain the viability of a mouse mast cell line in culture, but BCRFI lacks comparable activity in this way, suggesting that BCRFI may have conserved only a subset of hIL-10 activities.

Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: Homology to Epstein-Barr virus open reading frame BCRFI

International Nuclear Information System (INIS)

Vieira, P.; De Waal-Malefyt, R.; Dang, M.N.; Johnson, K.E.; Kastelein, R.; Fiorentino, D.F.; DeVries, J.E.; Roncarolo, M.G.; Mosmann, T.R.; Moore, K.W.

1991-01-01

The authors demonstrated the existence of human cytokine synthesis inhibitory factor (DSIF) [interleukin 10 (IL-10)]. cDNA clones encoding human IL-10 (hIL-10) were isolated from a tetanus toxin-specific human T-cell clone. Like mouse IL-10, hIL-10 exhibits strong DNA and amino acid sequence homology to an open reading frame in the Epstein-Barr virus, BDRFL. hIL-10 and the BCRFI product inhibit cytokine synthesis by activated human peripheral blood mononuclear cells and by a mouse Th1 clone. Both hIL-10 and mouse IL-10 sustain the viability of a mouse mast cell line in culture, but BCRFI lacks comparable activity in this way, suggesting that BCRFI may have conserved only a subset of hIL-10 activities

Salicylic acid alleviates the adverse effects of salt stress in Torreya grandis cv. Merrillii seedlings by activating photosynthesis and enhancing antioxidant systems.

Directory of Open Access Journals (Sweden)

Tingting Li

Full Text Available BACKGROUND: Salt stress is a major factor limiting plant growth and productivity. Salicylic acid (SA has been shown to ameliorate the adverse effects of environmental stress on plants. To investigate the protective role of SA in ameliorating salt stress on Torreya grandis (T. grandis trees, a pot experiment was conducted to analyze the biomass, relative water content (RWC, chlorophyll content, net photosynthesis (Pn, gas exchange parameters, relative leakage conductivity (REC, malondialdehyde (MDA content, and activities of superoxide dismutase (SOD and peroxidase (POD of T. grandis under 0.2% and 0.4% NaCl conditions with and without SA. METHODOLOGY/PRINCIPAL FINDINGS: The exposure of T. grandis seedlings to salt conditions resulted in reduced growth rates, which were associated with decreases in RWC and Pn and increases in REC and MDA content. The foliar application of SA effectively increased the chlorophyll (chl (a+b content, RWC, net CO2 assimilation rates (Pn, and proline content, enhanced the activities of SOD, CAT and POD, and minimized the increases in the REC and MDA content. These changes increased the capacity of T. grandis in acclimating to salt stress and thus increased the shoot and root dry matter. However, when the plants were under 0% and 0.2% NaCl stress, the dry mass of the shoots and roots did not differ significantly between SA-treated plants and control plants. CONCLUSIONS: SA induced the salt tolerance and increased the biomass of T. grandis cv. by enhancing the chlorophyll content and activity of antioxidative enzymes, activating the photosynthetic process, and alleviating membrane injury. A better understanding about the effect of salt stress in T. grandis is vital, in order gain knowledge over expanding the plantations to various regions and also for the recovery of T. grandis species in the future.

Isolation and characterization of a cDNA encoding phytochrome A in the non-photosynthetic parasitic plant, Orobanche minor Sm.

Science.gov (United States)

Trakulnaleamsai, Chitra; Okazawa, Atsushi; An, Chung-Il; Kajiyama, Shin'ichiro; Fukusaki, Ei'ichiro; Yoneyama, Koichi; Takeuchi, Yasutomo; Kobayashi, Akio

2005-01-01

In this study, the isolation and characterization of a phytochrome A (PHYA) homologous cDNA (OmPHYA) in the non-photosynthetic holoparasitic plant Orobanche minor are described. The present findings provide the first report of the presence of a PHYA homolog in the holoparasite. This study found that OmPHYA is of similar size to the other PHYAs of green plants and shows 72, 77, and 77% amino acid sequence identity with PHYA in Arabidopsis, potato, and tobacco respectively. The OmPHYA contains a conserved chromophore attachment cysteine at position 323. Although OmPHYA shows high sequence identity with other PHYAs in green plants, 13 amino acid substitutions located in both the N and C-terminal domains are observed (a total of 26 amino acids). OmPHYA is encoded by a single gene within the O. minor genome. The abundance of the OmPHYA transcript as well as nuclear translocation of OmphyA occurs in a light-dependent manner.

A Newly Identified Passive Hyperaccumulator Eucalyptus grandis × E. urophylla under Manganese Stress.

Directory of Open Access Journals (Sweden)

Qingqing Xie

Full Text Available Manganese (Mn is an essential micronutrient needed for plant growth and development, but can be toxic to plants in excess amounts. However, some plant species have detoxification mechanisms that allow them to accumulate Mn to levels that are normally toxic, a phenomenon known as hyperaccumulation. These species are excellent candidates for developing a cost-effective remediation strategy for Mn-polluted soils. In this study, we identified a new passive Mn-hyperaccumulator Eucalyptus grandis × E. urophylla during a field survey in southern China in July 2010. This hybrid can accumulate as much as 13,549 mg/kg DW Mn in its leaves. Our results from Scanning Electron Microscope (SEM X-ray microanalysis indicate that Mn is distributed in the entire leaf and stem cross-section, especially in photosynthetic palisade, spongy mesophyll tissue, and stem xylem vessels. Results from size-exclusion chromatography coupled with ICP-MS (Inductively coupled plasma mass spectrometry lead us to speculate that Mn associates with relatively high molecular weight proteins and low molecular weight organic acids, including tartaric acid, to avoid Mn toxicity. Our results provide experimental evidence that both proteins and organic acids play important roles in Mn detoxification in Eucalyptus grandis × E. urophylla. The key characteristics of Eucalyptus grandis × E. urophylla are an increased Mn translocation facilitated by transpiration through the xylem to the leaves and further distribution throughout the leaf tissues. Moreover, the Mn-speciation profile obtained for the first time in different cellular organelles of Eucalyptus grandis × E. urophylla suggested that different organelles have differential accumulating abilities and unique mechanisms for Mn-detoxification.

Hibiscus latent Fort Pierce virus in Brazil and synthesis of its biologically active full-length cDNA clone.

Science.gov (United States)

Gao, Ruimin; Niu, Shengniao; Dai, Weifang; Kitajima, Elliot; Wong, Sek-Man

2016-10-01

A Brazilian isolate of Hibiscus latent Fort Pierce virus (HLFPV-BR) was firstly found in a hibiscus plant in Limeira, SP, Brazil. RACE PCR was carried out to obtain the full-length sequences of HLFPV-BR which is 6453 nucleotides and has more than 99.15 % of complete genomic RNA nucleotide sequence identity with that of HLFPV Japanese isolate. The genomic structure of HLFPV-BR is similar to other tobamoviruses. It includes a 5' untranslated region (UTR), followed by open reading frames encoding for a 128-kDa protein and a 188-kDa readthrough protein, a 38-kDa movement protein, 18-kDa coat protein, and a 3' UTR. Interestingly, the unique feature of poly(A) tract is also found within its 3'-UTR. Furthermore, from the total RNA extracted from the local lesions of HLFPV-BR-infected Chenopodium quinoa leaves, a biologically active, full-length cDNA clone encompassing the genome of HLFPV-BR was amplified and placed adjacent to a T7 RNA polymerase promoter. The capped in vitro transcripts from the cloned cDNA were infectious when mechanically inoculated into C. quinoa and Nicotiana benthamiana plants. This is the first report of the presence of an isolate of HLFPV in Brazil and the successful synthesis of a biologically active HLFPV-BR full-length cDNA clone.

Human uroporphyrinogen III synthase: Molecular cloning, nucleotide sequence, and expression of a full-length cDNA

International Nuclear Information System (INIS)

Tsai, Shihfeng; Bishop, D.F.; Desnick, R.J.

1988-01-01

Uroporphyrinogen III synthase, the fourth enzyme in the heme biosynthetic pathway, is responsible for conversion of the linear tetrapyrrole, hydroxymethylbilane, to the cyclic tetrapyrrole, uroporphyrinogen III. The deficient activity of URO-synthase is the enzymatic defect in the autosomal recessive disorder congenital erythropoietic porphyria. To facilitate the isolation of a full-length cDNA for human URO-synthase, the human erythrocyte enzyme was purified to homogeneity and 81 nonoverlapping amino acids were determined by microsequencing the N terminus and four tryptic peptides. Two synthetic oligonucleotide mixtures were used to screen 1.2 x 10 6 recombinants from a human adult liver cDNA library. Eight clones were positive with both oligonucleotide mixtures. Of these, dideoxy sequencing of the 1.3 kilobase insert from clone pUROS-2 revealed 5' and 3' untranslated sequences of 196 and 284 base pairs, respectively, and an open reading frame of 798 base pairs encoding a protein of 265 amino acids with a predicted molecular mass of 28,607 Da. The isolation and expression of this full-length cDNA for human URO-synthase should facilitate studies of the structure, organization, and chromosomal localization of this heme biosynthetic gene as well as the characterization of the molecular lesions causing congenital erythropoietic porphyria

ECONOMIC ROTATION OF Eucalyptus grandis PLANTATIONS FOR PULP PRODUCTION

Directory of Open Access Journals (Sweden)

Thais Cunha Ferreira

2004-07-01

Full Text Available The objectives of the research were: to determine the economic impact of several minimum diameter and length of logs in economic rotation age, economic feasibility of Eucalyptus grandis plantation for cellulose production; to determine the economic loss of cutting the stand before or after the optimal economic rotation age. A biometric model for making wood volume prognosis was developed using data of a trial of Eucalyptus grandis plantation envisaging pulp production. Eucalyptus grandis stands of 19 and 103 months old, in the spacing 3 x 2 and 3 x 3 m in site index of 30; 28; 26 and 24 m were used. Theprognosis started at the age zero, considering logs of 2.5; 2.8; 4.0 and 6.0 m of length for minimum diameter varying from 4 to 10 cm, in intervals of 2 cm. Net Present Worth (VPL was used the economic criterion, considering an infinite horizon and a cost relation including reestablishment, yearly maintenance, logging and wood transportation costs. The main conclusions were: increases in the minimum diameter and or in logs length increase the rotation age; harvesting the stands in ages different from the optimal one cause large economic loss mainly in the better sites; the economic loss is larger if the harvest is made before the optimal economic rotation than if it is make after; economic feasibility increases when the minimum diameter is smaller and when the length of the logs is shorter. Any way, before making any decision it is necessary to take into account possible technical restrictions and effect on harvest and transportation costs caused by changer in the length of logs and in the size of the minimum commercial diameter.

Dry mass allocation, water use efficiency and delta C-13 in clones of Eucalyptus grandis, E-grandis x camaldulensis and E-grandis x nitens grown under two irrigation regimes

CSIR Research Space (South Africa)

Le Roux, D

1996-05-01

Full Text Available - cial clones of Eucalyptus grandis W. Hill ex Maiden. implying that less water-use-efficient trees were more productive (Bond and Stock 1990). Similarly, growing season WUE and delta13C were positively correlated in western larch and Eucalyptus globulus... regimes DEBBIE LE ROUX,1,2 WILLIAM D. STOCK,3 WILLIAM J. BOND3 and DAVID MAPHANGA4 1 Division of Forest Science and Technology, CSIR, Pretoria, South Africa 2 Present address: Department of Botany, University of Kansas, Lawrence, KS 66045, USA 3 Department...

Human tissue factor: cDNA sequence and chromosome localization of the gene

International Nuclear Information System (INIS)

Scarpati, E.M.; Wen, D.; Broze, G.J. Jr.; Miletich, J.P.; Flandermeyer, R.R.; Siegel, N.R.; Sadler, J.E.

1987-01-01

A human placenta cDNA library in λgt11 was screened for the expression of tissue factor antigens with rabbit polyclonal anti-human tissue factor immunoglobulin G. Among 4 million recombinant clones screened, one positive, λHTF8, expressed a protein that shared epitopes with authentic human brain tissue factor. The 1.1-kilobase cDNA insert of λHTF8 encoded a peptide that contained the amino-terminal protein sequence of human brain tissue factor. Northern blotting identified a major mRNA species of 2.2 kilobases and a minor species of ∼ 3.2 kilobases in poly(A) + RNA of placenta. Only 2.2-kilobase mRNA was detected in human brain and in the human monocytic U937 cell line. In U937 cells, the quantity of tissue factor mRNA was increased several fold by exposure of the cells to phorbol 12-myristate 13-acetate. Additional cDNA clones were selected by hybridization with the cDNA insert of λHTF8. These overlapping isolates span 2177 base pairs of the tissue factor cDNA sequence that includes a 5'-noncoding region of 75 base pairs, an open reading frame of 885 base pairs, a stop codon, a 3'-noncoding region of 1141 base pairs, and a poly(a) tail. The open reading frame encodes a 33-kilodalton protein of 295 amino acids. The predicted sequence includes a signal peptide of 32 or 34 amino acids, a probable extracellular factor VII binding domain of 217 or 219 amino acids, a transmembrane segment of 23 acids, and a cytoplasmic tail of 21 amino acids. There are three potential glycosylation sites with the sequence Asn-X-Thr/Ser. The 3'-noncoding region contains an inverted Alu family repetitive sequence. The tissue factor gene was localized to chromosome 1 by hybridization of the cDNA insert of λHTF8 to flow-sorted human chromosomes

[Distribution pattern of meso-micro soil fauna in Eucalyptus grandis plantation].

Science.gov (United States)

Huang, Yumei; Zhang, Jian; Yang, Wanqin

2006-12-01

In this paper, meso-micro soil fauna were extracted and collected by Baermann's and Tullgren' s method, and their distribution pattern in the Eucalyptus grandis plantation of Hongya County, Sichuan Province was studied. A total of 13 550 specimens were collected, belonging to 6 phyla, 13 classes, and 26 orders. Acarina, Nematoda, Collembola were the dominant groups, and Enchytraeidae was the frequent one. The group and individual numbers of meso-micro soil fauna varied with seasons, being the maximum in autumn or winter, fewer in summer, and the minimum in spring. The density of meso-micro soil fauna in soil profile decreased rapidly with increasing soil depth, but a converse distribution was observed from time to time in 5 - 10 cm and 10 - 15 cm soil layers. The meso-micro soil fauna collected by Baermann's and Tullgren's method had a density of 3. 333 x 10(3) - 2. 533 x 10(5) ind x m(-2) and 1.670 x 10(2) - 2.393 x 10(5) ind x m(-2), respectively, and the decreasing rate of the density with the increase of soil depth was higher for those collected by Tullgren's method. The density-group index of meso-micro soil fauna in the E. grandis plantation was the lowest in spring, but the highest in autumn or summer. There were no significant differences in the density of meso-micro soil fauna and in the density-group index between E. grandis plantation and Quercus acutissima secondary forest.

Isolation and characterization of the cDNA for cystic fibrosis antigen

Energy Technology Data Exchange (ETDEWEB)

Dorin, J R

1987-01-01

Cystic fibrosis antigen (CFAg) is a protein which is present in the serum of cystic fibrosis (CF) patients and clinically normal heterozygotes, but not in normal individuals. CFAg has been shown to be a major granulocyte protein in normals, and the gene mapped to chromosome 1. This thesis describes the molecular cloning and subsequent characterization of the cDNA for CFAg. The availability of monoclonal antibodies to CFAg facilitated the identification of myeloid tissues which were actively synthesizing the protein. A specific radiolabeled protein could be immunoprecipitated from /sup 35/S-methionine labelled extracts of chronic myeloid leukemia cells (CML), normal granulocytes and HL-60 cells differentiated towards granulocytes. In CML and granulocytes, CFAg appeared to be one of the most abundantly synthesized proteins.

The potential of young, green finger-jointed Eucalyptus grandis ...

African Journals Online (AJOL)

Drying will occur naturally while the lumber is fixed within the roof truss structure. The objectives of this study were (1) to investigate the strength and stiffness variation of the finger-jointed E. grandis product in both the green and dry state for different age and dimension lumber, (2) to investigate the variation in density, warp ...

A Multivariate Study on Genetic Variation in Teak (Tectona grandis (L.))

DEFF Research Database (Denmark)

Kjær, Erik Dahl; Siegismund, Hans Redlef; Suangtho, V.

1996-01-01

Genetic differentiation between populations of teak (Tectona grandis (L.)) was examined in 9 quantitative characters and 10 allozyme loci. Large differences between populations were revealed in the quantitative traits. Regional patterns were revealed by multivariate analysis of the data, but ther...

Establishment of Eucalyptus grandis W. Hill ex Maiden in vitro using ...

African Journals Online (AJOL)

Establishment of Eucalyptus grandis W. Hill ex Maiden in vitro using commercial products for seed treatment. Moacir Ribeiro Neto, Cíntia de Oliveira Martendal, Flávia Dionísio Pereira, Edson Luiz Souchie, Fabiano Guimarães Silva ...

The function analysis of full-length cDNA sequence from IRM-2 mouse cDNA library

International Nuclear Information System (INIS)

Wang Qin; Liu Xiaoqiu; Xu Chang; Du Liqing; Sun Zhijuan; Wang Yan; Liu Qiang; Song Li; Li Jin; Fan Feiyue

2013-01-01

Objective: To identify the function of full-length cDNA sequence from IRM-2 mouse cDNA library. Methods: Full-length cDNA products were amplified by PCR from IRM-2 mouse cDNA library according to twenty-one pieces of expressed sequence tag. The expression of full-length cDNAs were detected after mouse embryonic fibroblasts were exposed to 6.5 Gy γ-ray radiation. And the effect on the growth of radiosensitivity cells AT5B1VA transfected with full-length cDNAs was investigated. Results: The expression of No.4, 5 and 2 full-length cDNAs from IRM-2 mouse were higher than that of parental ICR and 615 mouse after mouse embryonic fibroblasts irradiated with γ-ray radiation. And the survival rate of AT5B1VA cells transfected with No.4, 5 and 2 full-length cDNAs was high. Conclusion: No.4, 5 and 2 full-length cDNAs of IRM-2 mouse are of high radioresistance. (authors)

Grandi Eventi: indicatori di classificazione e incidenza sui sistemi urbani

Directory of Open Access Journals (Sweden)

Giuseppe Mazzeo

2008-08-01

Full Text Available I grandi eventi sono ritenuti una opportunità per le città in quanto sono uno straordinario catalizzatore di investimenti mirati alla trasformazione della città. L’azione dei grandi eventi non è confinata solo nel periodo di svolgimento dell’evento stesso ma si estende ad un periodo molto più ampio, prima e, soprattutto, dopo il suo termine. Nell’organizzazione delle manifestazioni più recenti le trasformazioni urbane hanno interessato in modo sempre maggiore parti già in precedenza urbanizzate, al punto che l’evento diviene l’occasione per trasformare la città costruita e per modificarne il profilo. Il paper approfondisce l’argomento della classificazione degli eventi e del loro impatto sul sistema urbano sviluppandosi in tre parti. Nella prima si analizzano due tipologie di grandi eventi (le esposizioni internazionali e i giochi olimpici con l’obiettivo di pervenire ad una definizione condivisa. Nella seconda parte si identificano i più importanti fattori ed indicatori per l’analisi di questa specifica categoria di avvenimenti. Infine, nella terza parte, si approfondiscono i fattori connessi all’impatto sul sistema urbano e all’organizzazione della sua mobilità. In particolare, viene approfondito il concetto di “effetto pulsar”, ossia gli effetti moltiplicativi sulla evoluzione urbana dipendenti dalla organizzazione di eventi multipli in parallelo o in sequenza. Uno degli elementi più negativi connessi alla organizzazione di un grande evento è la dispersione nel tempo dei benefici acquisiti grazie ad esso; per evitare ciò è necessario che l’evento sia seguito da politiche strategiche continue in modo da preservare i vantaggi acquisiti.

Data on internal cDNA amplification and color changes of the proteins derived from Pacific white leg shrimp shell.

Science.gov (United States)

Chuang, Pan; Shoichiro, Ishizaki; Yuji, Nagashima; Jialong, Gao; Shugo, Watabe

2018-02-01

In this article, we report original data on the designation of the primers for full-length cDNA amplification and the internal cDNA amplification of red color-related pigment-binding protein derived from shrimp shell. Data on the color shifts of different soluble proteins under 100 °C 10 min heat treatment and the effects of heating temperatures (from 30 to 100 °C) on the color changes of crude water-soluble proteins are also included in this report. For further details and experimental findings please refer to the article "Isolation and cDNA cloning of a novel red color-related pigment-binding protein derived from the shell of shrimp, Litopenaeus vannamei " (Chuang et al., 2017) [1].

Data on internal cDNA amplification and color changes of the proteins derived from Pacific white leg shrimp shell

Directory of Open Access Journals (Sweden)

Pan Chuang

2018-02-01

Full Text Available In this article, we report original data on the designation of the primers for full-length cDNA amplification and the internal cDNA amplification of red color-related pigment-binding protein derived from shrimp shell. Data on the color shifts of different soluble proteins under 100 °C 10 min heat treatment and the effects of heating temperatures (from 30 to 100 °C on the color changes of crude water-soluble proteins are also included in this report. For further details and experimental findings please refer to the article “Isolation and cDNA cloning of a novel red color-related pigment-binding protein derived from the shell of shrimp, Litopenaeus vannamei” (Chuang et al., 2017 [1].

Dependence of rate of germination of teak ( Tectona grandis ) seeds ...

African Journals Online (AJOL)

A study was conducted to determine suitable sources of teak (Tectona grandis) seeds and methods of treating the seeds to promote higher rate of germination, with the objective to supply large quantities of seedlings for developing commercial teak plantations in Ghana. The field work involved seed collection, seed pericarp ...

Molecular cloning and expression of cDNA encoding a lumenal calcium binding glycoprotein from sarcoplasmic reticulum

International Nuclear Information System (INIS)

Leberer, E.; Charuk, J.H.M.; MacLennan, D.H.; Green, N.M.

1989-01-01

Antibody screening was used to isolate a cDNA encoding the 160-kDa glycoprotein of rabbit skeletal muscle sarcoplasmic reticulum. The cDNA is identical to that encoding the 53-kDa glycoprotein except that it contains an in-frame insertion of 1,308 nucleotides near its 5' end, apparently resulting from alternative splicing. The protein encoded by the cDNA would contain a 19-residue NH 2 -terminal signal sequence and a 453-residue COOH-terminal sequence identical to the 53-kDa glycoprotein. It would also contain a 436-amino acid insert between these sequences. This insert would be highly acidic, suggesting that it might bind Ca 2+ . The purified 160-kDa glycoprotein and the glycoprotein expressed in COS-1 cells transfected with cDNA encoding the 160-kDa glycoprotein were shown to bind 45 C 2+ in a gel overlay assay. The protein was shown to be located in the lumen of the sarcoplasmic reticulum and to be associated through Ca 2+ with the membrane. The authors propose that this lumenal Ca 2+ binding glycoprotein of the sarcoplasmic reticulum be designated sarcalumenin

cDNA cloning of human DNA topoisomerase I. Catalytic activity of a 67.7-kDa carboxyl-terminal fragment

International Nuclear Information System (INIS)

D'Arpa, P.; Machlin, P.S.; Ratrie, H. III; Rothfield, N.F.; Cleveland, D.W.; Earnshaw, W.C.

1988-01-01

cDNA clones encoding human topoisomerase I were isolated from an expression vector library (λgt11) screened with autoimmune anti-topoisomerase I serum. One of these clones has been expressed as a fusion protein comprised of a 32-kDa fragment of the bacterial TrpE protein linked to 67.7 kDa of protein encoded by the cDNA. Three lines of evidence indicate that the cloned cDNA encodes topoisomerase I. (i) Proteolysis maps of the fusion protein and human nuclear topoisomerase I are essentially identical. (ii) The fusion protein relaxes supercoiled DNA, an activity that can be immunoprecipitated by anti-topoisomerase I serum. (iii) Sequence analysis has revealed that the longest cDNA clone (3645 base pairs) encodes a protein of 765 amino acids that shares 42% identity with Saccharomyces cerevisiae topoisomerase I. The sequence data also show that the catalytically active 67.7-kDa fragment is comprised of the carboxyl terminus

Durabilidad natural de madera de Eucalyptus grandis Hill ex Maiden de plantaciones de rápido crecimiento

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Silvia Böthig

2011-05-01

Full Text Available Una de las especies forestales de rápido crecimiento cultivadas en Uruguay de mayor importancia económica es el Eucalyptus grandis. Trabajos anteriores reportan propiedades físicas y mecánicas de la madera juvenil y adulta proveniente de plantaciones de esta especie en diferentes regiones del país. Sin embargo, dado que no se dispone de datos científicos nacionales sobre su durabilidad natural, en este trabajo fue evaluada madera de E. grandis obtenida de dos plantaciones de 16 años de edad con semilla del mismo origen, de dos sitios, Rivera y Río Negro. Se estudió la durabilidad natural del duramen externo e interno siguiendo los métodos de la norma EN 350-1, tomando Populus deltoides x euroamericana cv I-214 como especie de referencia. Se realizaron ensayos de laboratorio para determinar la resistencia a la descomposición fúngica (Gloeophyllum trabeum, Trametes versicolor y Serpula lacrymans y a las termites (Reticulitermes spp. siguiendo las normas EN 113 y EN 118, respectivamente. Se realizaron ensayos de campo de estacas, de doble capa y cámara fúngica, los cuales aún están en curso, por lo que en el presente trabajo se presentan resultados parciales. Los perfiles radiales de densidad básica revelaron que la madera del duramen externo no era adulta, sino madera de transición. En general, el duramen de E. grandis mostró una mayor durabilidad que el híbrido Populus.El E. grandis se clasificó como moderadamente o seriamente atacado por Reticulitermes spp. Según EN 350-1, la madera juvenil de E. grandis puede considerarse como “moderadamente durable” ante la pudrición parda provocada por G. trabeum, mientras que la madera de transición puede describirse como “durable”. La madera de transición mostró en relación a la madera juvenil una mayor resistencia al G. trabeum, una susceptibilidad levemente menor a las termites y mejor desempeño en el campo luego de 17 meses de exposición. El sitio de la plantación no

Effect of thermal modification on the physical properties of juvenile and mature woods of Eucalyptus grandis

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Fred Willians Calonego

Full Text Available This study aimed to evaluate the effect of thermal treatment on the physical properties of juvenile and mature woods of Eucalyptus grandis. Boards were taken from 30-year-old E. grandis trees. The boards were thermally modified at 180 °C in the Laboratory of Wood Drying and Preservation at UNESP, Botucatu, Sao Paulo state, Brazil. The results showed that thermal modification caused: (1 decrease of 6.8% in the density at 0% equilibrium moisture content of mature wood; (2 significant decreases of 14.7% and 35.6% in the maximum volumetric swellings of juvenile and mature woods, respectively; (3 significant decreases of 13.7% and 21.3% in the equilibrium moisture content of juvenile and mature woods, respectively. The influence of thermal modification in juvenile wood was lower than in mature wood and caused greater uniformity in the physical variations between these types of wood in E. grandis.

Synthesis of 2,2,4,4-tetramethyl-N,N'-bis(2,6-dimethylphenyl)cyclobutane-1,3-diimine , a unique compound from Arundo donax, and its analogues to test their antifeedant activity against the boll weevil, Anthonomus grandis.

Science.gov (United States)

Mochizuki, K; Takikawa, H; Mori, K

2000-03-01

2,2,4,4-Tetramethyl-N,N'-bis(2,6-dimethylphenyl) cyclobutane-1,3-diimine (1), which was isolated from the Thai plant Arundo donax as an antifeedant against the boll weevil (Anthonomus grandis), and its analogues (9-13) were synthesized and shown to possess no remarkable antifeedant activity of practical interest.

Development and assessment of microarray-based DNA fingerprinting in Eucalyptus grandis.

Science.gov (United States)

Lezar, Sabine; Myburg, A A; Berger, D K; Wingfield, M J; Wingfield, B D

2004-11-01

Development of improved Eucalyptus genotypes involves the routine identification of breeding stock and superior clones. Currently, microsatellites and random amplified polymorphic DNA markers are the most widely used DNA-based techniques for fingerprinting of these trees. While these techniques have provided rapid and powerful fingerprinting assays, they are constrained by their reliance on gel or capillary electrophoresis, and therefore, relatively low throughput of fragment analysis. In contrast, recently developed microarray technology holds the promise of parallel analysis of thousands of markers in plant genomes. The aim of this study was to develop a DNA fingerprinting chip for Eucalyptus grandis and to investigate its usefulness for fingerprinting of eucalypt trees. A prototype chip was prepared using a partial genomic library from total genomic DNA of 23 E. grandis trees, of which 22 were full siblings. A total of 384 cloned genomic fragments were individually amplified and arrayed onto glass slides. DNA fingerprints were obtained for 17 individuals by hybridizing labeled genome representations of the individual trees to the 384-element chip. Polymorphic DNA fragments were identified by evaluating the binary distribution of their background-corrected signal intensities across full-sib individuals. Among 384 DNA fragments on the chip, 104 (27%) were found to be polymorphic. Hybridization of these polymorphic fragments was highly repeatable (R2>0.91) within the E. grandis individuals, and they allowed us to identify all 17 full-sib individuals. Our results suggest that DNA microarrays can be used to effectively fingerprint large numbers of closely related Eucalyptus trees.

Comparisons between two economically valuable forest species (Eucalyptus grandis and Pinus taeda in relation to seed behaviour under controlled deterioration

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Jussara Bertho Fantinatti

2011-02-01

Full Text Available The objectives of this work were to analyze seed behaviour under controlled deterioration and estimate viability equations for forest species Eucalyptus grandis and Pinus taeda. Desired moisture content levels were achieved from initial values after either rehydration over water or drying over silica gel, both at 25 ºC. Seed sub samples with 8 moisture contents each for E. grandis (1.2 to 18.1%, initial value of 11.3% and P. taeda (1.5 to 19.5%, initial value of 12.9% were sealed in laminate aluminium-foil packets and stored in incubators maintained at 40, 50 and 65 ºC. The seeds from these species exhibited true orthodox and sub-orthodox storage behaviour, respectively, however E. grandis showed higher seed storability, probably due to a different seed chemical composition. Lowest moisture content limits estimated for application of the viability equations at 65 ºC were 4.9 and 4.1 mc for E. grandis and P. taeda, on equilibrium with ±20% RH. The viability equation estimated quantified the response of seed longevity to storage environment well with K E = 9.661 and 8.838; C W = 6.467 and 5.981; C H = 0.03498 and 0.10340; C Q = 0.0002330 and 0.0005476, for E. grandis and P. taeda, respectively.

ISOLASI cDNA SUCROSE TRANSPORTER (SUT DARI BATANG TANAMAN TEBU (Saccharum officinarum L.

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- Slameto

2010-09-01

Full Text Available Sucrose Transporter (SUT is kind of protein transporter that control in sucrose translocation. Sucrose Transporter is intermediate in translocation of sucrose from apoplasmic to simplasmic. SUT facilitates sucrose transportation from vascular tissues to parenchyma cells toward in node sugarcane stem. This research was purposed to isolate cDNA SUT from sugarcane stem, and cloned in Escherichia coli strain DH5α. Total RNA of sugarcane stem was isolated by single step method, then add with oligo dT in order to obtain the first strand of SUT cDNA then used as template for PCR. The primer used for PCR is 5’ –ggg ctg att gtg gcc atg tc- ‘3 (SUT-F and 5’ –tgc cct ttg tct ccg gaa cc- ‘3 (SUT-R. PCR was programmed as follow denaturation at 94°C for 2 minutes and 30 second, annealing at 54°C for 30 s, extension at 72°C 2 min and 7 min, and storage at 4°C for unlimited, It was for 30 cycles. Complementary DNA SUT from PCR ligalized to pTOPO bunt-end, then it cloned in to E. coli strain DH5α. The cloning resulted then be sequenced in order to observe the homologues with other nucleotides sequences of some plant using BLASTn program in GENE BANK NCBI and the level of homology determined by Genetyx program. The concentrated of total RNA isolated was 5,024 μg/μl, with purity of 1,85. Complementary DNA SUT fragment from PCR with size 2037 bp appropriated to the both of primer was used. Complementary DNA SUT fragment showed by analyzed some of restriction enzyme e.g. EcoRI, PstI and BamHI. Homologues of this cDNA SUT fragment was 100% to SoSUT 2A of sugarcane stem and 84% to OsSUT of rice plant (Casu et al ., 2003.

Construction of cDNA libraries from Pseudocercospora fijiensis Morelet infected leaves of the cultivars Calcutta 4 and Niyarma Yik

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Milady Mendoza-Rodríguez

2004-01-01

Full Text Available Molecular studies of plant-pathogen interaction are very important for the identification of gene (s related with the pathogenic process, as well as with the plant resistance. These gene (s could be use for the genetic improvement programs in order to obtain resistant cultivars. The aim of this work was to construct complementary DNA (cDNA libraries from infected leaves with Pseudocercospora fijiensis CCIBP-Pf1 isolated of two banana cultivars (a resistant one Calcutta4 and another one susceptible Niyarma Yik. First-strand cDNA synthesis, was made beginning with one microgram of total RNA by using oligo dT primer and cDNA quality was checked by Polimerase chain reaction (PCR with cytochrome b specific primers. Second-strand cDNA synthesis was performed by using the homopolymeric tailing with dC-BamH I + dT-Not I primer combination. Four cDNA libraries of infected plants at different times of infection with the pathogen were obtained. Forty one clones of one of the libraries of Niyarma Yik were sequenced and the obtained sequences correspond with genes related to fungi. Key words: Banana-Mycosphaerella fijiensis interaction,Black Sigatoka, Musa spp.

Cellulose nanocrystal from pomelo (C. Grandis osbeck) albedo: Chemical, morphology and crystallinity evaluation

Energy Technology Data Exchange (ETDEWEB)

Zain, Nor Fazelin Mat; Yusop, Salma Mohamad [Food Science Program, School of Chemical Sciences and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Selangor (Malaysia); Ahmad, Ishak [Polymer Research Centre (PORCE), School of Chemical Sciences and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, Selangor (Malaysia)

2013-11-27

Citrus peel is one of the under-utilized waste materials that have potential in producing a valuable fibre, which are cellulose and cellulose nanocrystal. Cellulose was first isolated from pomelo (C. Grandis Osbeck) albedo by combination of alkali treatment and bleaching process, followed by acid hydrolysis (65% H{sub 2}SO{sub 4}, 45 °C, 45min) to produce cellulose nanocrystal. The crystalline, structural, morphological and chemical properties of both materials were studied. Result reveals the crystallinity index obtained from X-ray diffraction for cellulose nanocrystal was found higher than extracted cellulose with the value of 60.27% and 57.47%, respectively. Fourier transform infrared showed that the chemical treatments removed most of the hemicellulose and lignin from the pomelo albedo fibre. This has been confirmed further by SEM and TEM for their morphological studies. These results showed that cellulose and cellulose nanocrystal were successfully obtained from pomelo albedo and might be potentially used in producing functional fibres for food application.

Cellulose nanocrystal from pomelo (C. Grandis osbeck) albedo: Chemical, morphology and crystallinity evaluation

International Nuclear Information System (INIS)

Zain, Nor Fazelin Mat; Yusop, Salma Mohamad; Ahmad, Ishak

2013-01-01

Citrus peel is one of the under-utilized waste materials that have potential in producing a valuable fibre, which are cellulose and cellulose nanocrystal. Cellulose was first isolated from pomelo (C. Grandis Osbeck) albedo by combination of alkali treatment and bleaching process, followed by acid hydrolysis (65% H 2 SO 4 , 45 °C, 45min) to produce cellulose nanocrystal. The crystalline, structural, morphological and chemical properties of both materials were studied. Result reveals the crystallinity index obtained from X-ray diffraction for cellulose nanocrystal was found higher than extracted cellulose with the value of 60.27% and 57.47%, respectively. Fourier transform infrared showed that the chemical treatments removed most of the hemicellulose and lignin from the pomelo albedo fibre. This has been confirmed further by SEM and TEM for their morphological studies. These results showed that cellulose and cellulose nanocrystal were successfully obtained from pomelo albedo and might be potentially used in producing functional fibres for food application

Infectious Maize rayado fino virus from Cloned cDNA.

Science.gov (United States)

Edwards, Michael C; Weiland, John J; Todd, Jane; Stewart, Lucy R

2015-06-01

A full-length cDNA clone was produced from a U.S. isolate of Maize rayado fino virus (MRFV), the type member of the genus Marafivirus within the family Tymoviridae. Infectivity of transcripts derived from cDNA clones was demonstrated by infection of maize plants and protoplasts, as well as by transmission via the known leafhopper vectors Dalbulus maidis and Graminella nigrifrons that transmit the virus in a persistent-propagative manner. Infection of maize plants through vascular puncture inoculation of seed with transcript RNA resulted in the induction of fine stipple stripe symptoms typical of those produced by wild-type MRFV and a frequency of infection comparable with that of the wild type. Northern and Western blotting confirmed the production of MRFV-specific RNAs and proteins in infected plants and protoplasts. An unanticipated increase in subgenomic RNA synthesis over levels in infected plants was observed in protoplasts infected with either wild-type or cloned virus. A conserved cleavage site motif previously demonstrated to function in both Oat blue dwarf virus capsid protein and tymoviral nonstructural protein processing was identified near the amino terminus of the MRFV replicase polyprotein, suggesting that cleavage at this site also may occur.

Effects of autohydrolysis of Eucalyptus urograndis and Eucalyptus grandis on influence of chemical components and crystallinity index.

Science.gov (United States)

da Silva Morais, Alaine Patrícia; Sansígolo, Cláudio Angeli; de Oliveira Neto, Mario

2016-08-01

Samples of Eucalyptus urograndis and Eucalyptus grandis sawdust were autohydrolyzed in aqueous conditions to reach temperatures in the range 110-190°C and reaction times of 0-150min in a minireactor. In each minireactor were used a liquor:wood ratio (10:1 L:kg dry wood), in order to assess the effects of the autohydrolysis severity and the crystalline properties of cellulose. The content of extractives, lignin, holocellulose, cellulose, hemicelluloses and crystallinity index obtained from the solid fraction after autohydrolysis of sawdust were determined. This study demonstrated that the hemicelluloses were extensively removed at 170 and 190°C, whereas cellulose was partly degraded to Eucalyptus urograndis and Eucalyptus grandis sawdust. The lignin content decreased, while the extractives content increased. It was defined that during autohydrolysis, had a slight decreased on crystalline structure of cellulose of Eucalyptus urogandis and Eucalyptus grandis. Copyright © 2016 Elsevier Ltd. All rights reserved.

Human pro. cap alpha. 1(III) collagen: cDNA sequence for the 3' end

Energy Technology Data Exchange (ETDEWEB)

Mankoo, B S; Dalgleish, R

1988-03-25

The authors have previously isolated two overlapping cDNA clones, pIII-21 and pIII-33, which encode the C-terminal end of human type III procollagen. They now present the sequence of 2520 bases encoded in these cDNAs which overlaps other previously published sequences for the same gene. The sequence presented differs from previously published sequences at five positions.

Leaf cDNA-AFLP analysis reveals novel mechanisms for boron-induced alleviation of aluminum-toxicity in Citrus grandis seedlings.

Science.gov (United States)

Wang, Liu-Qing; Yang, Lin-Tong; Guo, Peng; Zhou, Xin-Xing; Ye, Xin; Chen, En-Jun; Chen, Li-Song

2015-10-01

Little information is available on the molecular mechanisms of boron (B)-induced alleviation of aluminum (Al)-toxicity. 'Sour pummelo' (Citrus grandis) seedlings were irrigated for 18 weeks with nutrient solution containing different concentrations of B (2.5 or 20μM H3BO3) and Al (0 or 1.2mM AlCl3·6H2O). B alleviated Al-induced inhibition in plant growth accompanied by lower leaf Al. We used cDNA-AFLP to isolate 127 differentially expressed genes from leaves subjected to B and Al interactions. These genes were related to signal transduction, transport, cell wall modification, carbohydrate and energy metabolism, nucleic acid metabolism, amino acid and protein metabolism, lipid metabolism and stress responses. The ameliorative mechanisms of B on Al-toxicity might be related to: (a) triggering multiple signal transduction pathways; (b) improving the expression levels of genes related to transport; (c) activating genes involved in energy production; and (d) increasing amino acid accumulation and protein degradation. Also, genes involved in nucleic acid metabolism, cell wall modification and stress responses might play a role in B-induced alleviation of Al-toxicity. To conclude, our findings reveal some novel mechanisms on B-induced alleviation of Al-toxicity at the transcriptional level in C. grandis leaves. Copyright © 2015 Elsevier Inc. All rights reserved.

Purification, cDNA Cloning, and Developmental Expression of the Nodule-Specific Uricase from Phaseolus vulgaris L. 1

Science.gov (United States)

Sánchez, Federico; Campos, Francisco; Padilla, Jaime; Bonneville, Jean-Marc; Enríquez, Consuelo; Caput, Daniel

1987-01-01

Nodule-specific uricase (uricase II) from Phaseolus vulgaris L. was purified to homogeneity by chromatographic methods. Purification data indicated that uricase II is approximately 2% of the total soluble protein from mature nodules. Specific antiserum was raised and used to determine the developmental expression and for immunoselection of polysomes. Uricase II was antigenically detected early in nodule development, 2 to 3 days before nitrogen fixation. Uricase-encoding cDNA clones were isolated by hybridizing a nodule-specific pUC9 cDNA library with labeled mRNA from immunoselected polysomes and a 35,000 molecular weight uricase II-encoding cDNA from soybean. An homologous clone (pNF-UR07) was used to assess the expression pattern of the specific transcript during development. Northern-blot analysis indicated that uricase II mRNA is exclusively expressed in nodule tissue. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:16665575

Second-strand cDNA synthesis: classical method

International Nuclear Information System (INIS)

Gubler, U.

1987-01-01

The classical scheme for the synthesis of double-stranded cDNA as it was reported in 1976 is described. Reverse transcription of mRNA with oligo(dT) as the primer generates first strands with a small loop at the 3' end of the cDNA (the end that corresponds to the 5' end of the mRNA). Subsequent removal of the mRNA by alkaline hydrolysis leaves single-stranded cDNA molecules again with a small 3' loop. This loop can be used by either reverse transcriptase or Klenow fragment of DNA polymerase I as a primer for second-strand synthesis. The resulting products are double-stranded cDNA molecules that are covalently closed at the end corresponding to the 5' end of the original mRNA. Subsequent cleavage of the short piece of single-stranded cDNA within the loop with the single-strand-specific S 1 nuclease generate open double-stranded molecules that can be used for molecular cloning in plasmids or in phage. Useful variations of this scheme have been described

Antraquinonas e naftoquinonas do caule de um espécime de reflorestamento de Tectona grandi (Verbenaceae)

OpenAIRE

Moreira,Rafael Y.O.; Arruda,Mara S.P.; Arruda,Alberto C.; Santos,Lourivaldo S.; Müller,Adolfo H.; Guilhon,Giselle M.S.P.; Santos,Alberdan S.; Terezo,Evaristo

2006-01-01

O fracionamento do extrato hexânico do caule de um espécime de reflorestamento de Tectona grandis (Verbenaceae), através de procedimentos fitoquímicos clássicos, levou ao isolamento das naftoquinonas lapachol e desidro-a-lapachona e das antraquinonas tectoquinona e obtusifolina. As estruturas das substâncias foram caracterizadas através da análise de métodos espectrométricos de RMN. Este é o primeiro estudo fitoquímico de um espécime de reflorestamento de Tectona grandis, no Brasil, sendo o o...

Preparation of a differentially expressed, full-length cDNA expression library by RecA-mediated triple-strand formation with subtractively enriched cDNA fragments

NARCIS (Netherlands)

Hakvoort, T. B.; Spijkers, J. A.; Vermeulen, J. L.; Lamers, W. H.

1996-01-01

We have developed a fast and general method to obtain an enriched, full-length cDNA expression library with subtractively enriched cDNA fragments. The procedure relies on RecA-mediated triple-helix formation of single-stranded cDNA fragments with a double-stranded cDNA plasmid library. The complexes

Improving Cry8Ka toxin activity towards the cotton boll weevil (Anthonomus grandis).

Science.gov (United States)

Oliveira, Gustavo R; Silva, Maria C M; Lucena, Wagner A; Nakasu, Erich Y T; Firmino, Alexandre A P; Beneventi, Magda A; Souza, Djair S L; Gomes, José E; de Souza, José D A; Rigden, Daniel J; Ramos, Hudson B; Soccol, Carlos R; Grossi-de-Sa, Maria F

2011-09-09

The cotton boll weevil (Anthonomus grandis) is a serious insect-pest in the Americas, particularly in Brazil. The use of chemical or biological insect control is not effective against the cotton boll weevil because of its endophytic life style. Therefore, the use of biotechnological tools to produce insect-resistant transgenic plants represents an important strategy to reduce the damage to cotton plants caused by the boll weevil. The present study focuses on the identification of novel molecules that show improved toxicity against the cotton boll weevil. In vitro directed molecular evolution through DNA shuffling and phage display screening was applied to enhance the insecticidal activity of variants of the Cry8Ka1 protein of Bacillus thuringiensis. Bioassays carried out with A. grandis larvae revealed that the LC50 of the screened mutant Cry8Ka5 toxin was 3.15-fold higher than the wild-type Cry8Ka1 toxin. Homology modelling of Cry8Ka1 and the Cry8Ka5 mutant suggested that both proteins retained the typical three-domain Cry family structure. The mutated residues were located mostly in loops and appeared unlikely to interfere with molecular stability. The improved toxicity of the Cry8Ka5 mutant obtained in this study will allow the generation of a transgenic cotton event with improved potential to control A. grandis.

Improving Cry8Ka toxin activity towards the cotton boll weevil (Anthonomus grandis

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Gomes José E

2011-09-01

Full Text Available Abstract Background The cotton boll weevil (Anthonomus grandis is a serious insect-pest in the Americas, particularly in Brazil. The use of chemical or biological insect control is not effective against the cotton boll weevil because of its endophytic life style. Therefore, the use of biotechnological tools to produce insect-resistant transgenic plants represents an important strategy to reduce the damage to cotton plants caused by the boll weevil. The present study focuses on the identification of novel molecules that show improved toxicity against the cotton boll weevil. In vitro directed molecular evolution through DNA shuffling and phage display screening was applied to enhance the insecticidal activity of variants of the Cry8Ka1 protein of Bacillus thuringiensis. Results Bioassays carried out with A. grandis larvae revealed that the LC50 of the screened mutant Cry8Ka5 toxin was 3.15-fold higher than the wild-type Cry8Ka1 toxin. Homology modelling of Cry8Ka1 and the Cry8Ka5 mutant suggested that both proteins retained the typical three-domain Cry family structure. The mutated residues were located mostly in loops and appeared unlikely to interfere with molecular stability. Conclusions The improved toxicity of the Cry8Ka5 mutant obtained in this study will allow the generation of a transgenic cotton event with improved potential to control A. grandis.

Primary culture of cat intestinal epithelial cells in vitro and the cDNA library construction.

Science.gov (United States)

Zhao, Gui Hua; Liu, Ye; Cheng, Yun Tang; Zhao, Qing Song; Qiu, Xiao; Xu, Chao; Xiao, Ting; Zhu, Song; Liu, Gong Zhen; Yin, Kun

2018-06-26

Felids are the only definitive hosts of Toxoplasma gondii. To lay a foundation for screening the T. gondii-felids interaction factors, we have developed a reproducible primary culture method for cat intestinal epithelial cells (IECs). The primary IECs were isolated from a new born cat's small intestine jejunum region without food ingress, and respectively in vitro cultured by tissue cultivation and combined digestion method with collagenase XI and dispase I, then purified by trypsinization. After identification, the ds cDNA of cat IECs was synthesized for constructing pGADT7 homogenization three-frame plasmid, and transformed into the yeast Y187 for generating the cDNA library. Our results indicated that cultivation of primary cat IECs relays on combined digestion to form polarized and confluent monolayers within 3 days with typical features of normal epithelial cells. The purified cells cultured by digestion method were identified to be nature intestinal epithelial cells using immunohistochemical analysis and were able to maintain viability for at least 15 passages. The homogenizable ds cDNA, which is synthesized from the total RNA extracted from our cultured IECs, distributed among 0.5-2.0 kb, and generated satisfying three-frame cDNA library with the capacity of 1.2 × 106 and the titer of 5.2 × 107 pfu/mL. Our results established an optimal method for the culturing and passage of cat IECs model in vitro, and laid a cDNA library foundation for the subsequent interaction factors screening by yeast two-hybrid.

Tolerance of Anadenanthera peregrina to Eucalyptus camaldulensis and Eucalyptus grandis essential oil as condition for mixed plantation

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Neimar de Freitas Duarte

2012-06-01

Full Text Available With the purpose of selecting the species of woody Caatinga for mixed plantations with Eucalyptus spp., the allelophatic effects of E. camaldulensis and E. grandis essential oil were studied on the growth activities of Anadenanthera peregrina. The plants were closed in glass chambers in the presence of volatile oil of E. camaldulensis or E. grandis at the concentration of 13 nl.cm-3. The number of leaves, height and diameter at soil lever were compared before, immediately after and after 30 days. Chlorophyll a and b, carotenoids and dry mass were evaluated after the treatment application. There was no inhibitory effect of E. camaldulensis and E. grandis oils on A. peregrina. E. camaldulensis, which was more adapted to semi-arid conditions, was planted in mixture stands with two native legume species, inoculated with Rhizobium and arbuscular mycorrhizal fungi. E. camaldulensis did not inhibit native species growth after two years of cultivation.

MANEJO DO SOLO E CRESCIMENTO INICIAL DE Eucalyptus grandis Hill ex Maiden EM ARGISSOLO

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Juliana Prevedello

2013-01-01

Full Text Available The forest species cultivation with rapid growth in Brazil has increased, mainly due to the diverse use of its wood and climate adaptation. The cultivation with minimum tillage in the forest sector stands out a way to increase productivity, combined with the maintenance of biodiversity and soil conservation. This study was conducted at the experimental area of State Foundation for Agricultural Research - Research Center for Forestry in Santa Maria - RS, with aimed to evaluate the effect of the soil tillage methods on soil physical properties and on initial development of Eucalyptus grandis, in a sandy loam Typic Hapludalf. Four soil management practices were compared: no-tillage; chisel tillage; chisel tillage plus harrowing and; rotary tillage (rotary tiller, installed in a randomized block design with three replications. The soil under no-tillage conditioned lower initial growth of eucalyptus due higher soil penetration resistance and bulk density, when compared with treatments with mobilization. The root distribution analysis in soil, despite being a qualitative method, was effective in demonstrating the effect of soil tillage for the Eucalyptus grandis plantation. The soil tillage with mobilization resulted in a higher initial growth of Eucalyptus grandis. The chisel tillage effects in the soil physical properties persisted after one year of soil tillage.

Evaluation of Chemical Composition and Biological Activities of Citrus pseudolimon and Citrus grandis Peel Essential Oils

International Nuclear Information System (INIS)

Sajid, A.; Hanif, M.A.; Shahid, M.

2016-01-01

Essential oils and their volatile constituents are used extensively to prevent and treat human diseases. In the past decades, worldwide demand for citrus essential oils has greatly increased. Citrus essential oils containing 85-99 percent volatile and 1-15 percent non-volatile components. Essential oils from Citrus pseudolimon and Citrus grandis peels were extracted through steam distillation and characterized by GC-MS. C. pseudolimon has thirty six and C. grandis has thirty three total components; limonene 47.07 percent and 71.48 percent was the major component in both oils respectively. Antioxidant activity was checked by 2, 2-diphenyl-1-picrylhydrazyl radical assay and β-carotene/linoleic acid bleaching test. Both oils have modest activity. The antimicrobial potential was assessed against different bacterial and fungus strains. C. pseudolimon oil possessed strong activity against all tested strains while C. grandis has moderate activity. The antitumor activity was evaluated by potato disc assay, C. pseudolimon showed 81.25 inhibition. Hence the essential oils could have a great potential in pharmaceutical industry. (author)

Gas Exchange Characteristics in Tectona grandis L. Clones under Varying Concentrations of CO2 Levels

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S. Saravanan

2014-08-01

Full Text Available The Institute of Forest Genetics and Tree Breeding, Coimbatore, India functioning under the Indian Council of Forestry Research and Education, Dehara Dun, has a long term systematic tree improvement program for Tectona grandis aimed to enhancing productivity and screening of clones for site specific. In the process, twenty clones of T. grandis L. were studied for the physiological parameters and water use efficiency with reference to the elevated CO2 levels. CO2 enrichment studies in special chambers help in understanding the changes at individual level, and also at physiological, biochemical and genetic level. It also provides valuable information for establishing plantations at different geographic locations. Considerable variations were observed when the selected 20 clones of T. grandis were subjected to physiological studies under elevated CO2 conditions (600 and 900 mol mol-1. Eight clones exhibited superior growth coupled with favorable physiological characteristics including high photosynthetic rate, carboxylation and water use efficiency under elevated CO2 levels. Clones with minimal variation in physiological characteristics under elevated levels of CO2 suggest their ability to overcome physiological stresses and adapt to varying climatic conditions.

Method for construction of normalized cDNA libraries

Science.gov (United States)

Soares, Marcelo B.; Efstratiadis, Argiris

1998-01-01

This invention provides a method to normalize a directional cDNA library constructed in a vector that allows propagation in single-stranded circle form comprising: (a) propagating the directional cDNA library in single-stranded circles; (b) generating fragments complementary to the 3' noncoding sequence of the single-stranded circles in the library to produce partial duplexes; (c) purifying the partial duplexes; (d) melting and reassociating the purified partial duplexes to appropriate Cot; and (e) purifying the unassociated single-stranded circles, thereby generating a normalized cDNA library. This invention also provides normalized cDNA libraries generated by the above-described method and uses of the generated libraries.

Differential representation of sunflower ESTs in enriched organ-specific cDNA libraries in a small scale sequencing project

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Heinz Ruth A

2003-09-01

Full Text Available Abstract Background Subtractive hybridization methods are valuable tools for identifying differentially regulated genes in a given tissue avoiding redundant sequencing of clones representing the same expressed genes, maximizing detection of low abundant transcripts and thus, affecting the efficiency and cost effectiveness of small scale cDNA sequencing projects aimed to the specific identification of useful genes for breeding purposes. The objective of this work is to evaluate alternative strategies to high-throughput sequencing projects for the identification of novel genes differentially expressed in sunflower as a source of organ-specific genetic markers that can be functionally associated to important traits. Results Differential organ-specific ESTs were generated from leaf, stem, root and flower bud at two developmental stages (R1 and R4. The use of different sources of RNA as tester and driver cDNA for the construction of differential libraries was evaluated as a tool for detection of rare or low abundant transcripts. Organ-specificity ranged from 75 to 100% of non-redundant sequences in the different cDNA libraries. Sequence redundancy varied according to the target and driver cDNA used in each case. The R4 flower cDNA library was the less redundant library with 62% of unique sequences. Out of a total of 919 sequences that were edited and annotated, 318 were non-redundant sequences. Comparison against sequences in public databases showed that 60% of non-redundant sequences showed significant similarity to known sequences. The number of predicted novel genes varied among the different cDNA libraries, ranging from 56% in the R4 flower to 16 % in the R1 flower bud library. Comparison with sunflower ESTs on public databases showed that 197 of non-redundant sequences (60% did not exhibit significant similarity to previously reported sunflower ESTs. This approach helped to successfully isolate a significant number of new reported sequences

A putative peroxidase cDNA from turnip and analysis of the encoded protein sequence.

Science.gov (United States)

Romero-Gómez, S; Duarte-Vázquez, M A; García-Almendárez, B E; Mayorga-Martínez, L; Cervantes-Avilés, O; Regalado, C

2008-12-01

A putative peroxidase cDNA was isolated from turnip roots (Brassica napus L. var. purple top white globe) by reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). Total RNA extracted from mature turnip roots was used as a template for RT-PCR, using a degenerated primer designed to amplify the highly conserved distal motif of plant peroxidases. The resulting partial sequence was used to design the rest of the specific primers for 5' and 3' RACE. Two cDNA fragments were purified, sequenced, and aligned with the partial sequence from RT-PCR, and a complete overlapping sequence was obtained and labeled as BbPA (Genbank Accession No. AY423440, named as podC). The full length cDNA is 1167bp long and contains a 1077bp open reading frame (ORF) encoding a 358 deduced amino acid peroxidase polypeptide. The putative peroxidase (BnPA) showed a calculated Mr of 34kDa, and isoelectric point (pI) of 4.5, with no significant identity with other reported turnip peroxidases. Sequence alignment showed that only three peroxidases have a significant identity with BnPA namely AtP29a (84%), and AtPA2 (81%) from Arabidopsis thaliana, and HRPA2 (82%) from horseradish (Armoracia rusticana). Work is in progress to clone this gene into an adequate host to study the specific role and possible biotechnological applications of this alternative peroxidase source.

Effects of black-eyed pea trypsin/chymotrypsin inhibitor on proteolytic activity and on development of Anthonomus grandis.

Science.gov (United States)

Franco, Octávio L; dos Santos, Roseane C; Batista, João A N; Mendes, Ana Cristina M; de Araújo, Marcus Aurélio M; Monnerat, Rose G; Grossi-de-Sá, Maria Fátima; de Freitas, Sonia M

2003-06-01

The cotton boll weevil Anthonomus grandis (Boheman) is one of the major pests of cotton (Gossypium hirsutum L.) in tropical and sub-tropical areas of the New World. This feeds on cotton floral fruits and buds causing severe crop losses. Digestion in the boll weevil is facilitated by high levels of serine proteinases, which are responsible for the almost all proteolytic activity. Aiming to reduce the proteolytic activity, the inhibitory effects of black-eyed pea trypsin/chymotrypsin inhibitor (BTCI), towards trypsin and chymotrypsin from bovine pancreas and from midguts of A. grandis larvae and adult insects were analyzed. BTCI, purified from Vigna unguiculata (L.) seeds, was highly active against different trypsin-like proteinases studied and moderately active against the digestive chymotrypsin of adult insects. Nevertheless, no inhibitory activity was observed against chymotrypsin from A. grandis larval guts. To test the BTCI efficiency in vivo, neonate larvae were reared on artificial diet containing BTCI at 10, 50 and 100 microM. A reduction of larval weight of up to approximately 54% at the highest BTCI concentration was observed. At this concentration, the insect mortality was 65%. This work constitutes the first observation of a Bowman-Birk type inhibitor active in vitro and in vivo toward the cotton boll weevil A. grandis. The results of bioassays strongly suggest that BTCI may have potential as a transgene protein for use in engineered crop plants modified for heightened resistance to the cotton boll weevil.

Mycorrhizal symbionts of Pisonia grandis and P. sechellarum in Seychelles: identification of mycorrhizal fungi and description of new Tomentella species.

Science.gov (United States)

Suvi, Triin; Tedersoo, Leho; Abarenkov, Kessy; Beaver, Katy; Gerlach, Justin; Kõljalg, Urmas

2010-01-01

Nyctaginaceae includes species that are predominantly non-mycorrhizal or form arbuscular or ectomycorrhiza. Root-associated fungi were studied from P. grandis and P. sechellarum roots collected respectively on the islands of Cousin and Silhouette in Seychelles. In addition fungal sporocarps were collected from the sampling area. Fungal symbionts were identified from the roots by anatomotyping and rDNA sequencing; sporocarps collected were examined microscopically and sequenced. Three distantly related ectomycorrhizal fungal species belonging to Thelephoraceae were identified from the roots of P. grandis. Sporocarps also were found for two symbionts and described as new Tomentella species. In addition Tomentella species collected from other Seychelles islands were studied and described as new species if there was no close resemblance to previously established species. P. sechellarum was determined to be an arbuscular mycorrhizal plant; three arbuscular mycorrhizal fungal species were detected from the roots. P. grandis is probably associated only with species of Thelephoraceae throughout its area. Only five Tomentella species are known to form ectomycorrhiza with P. grandis and they never have been found to be associated with another host, suggesting adaptation of these fungi to extreme environmental conditions in host's habitat.

Horse cDNA clones encoding two MHC class I genes

Energy Technology Data Exchange (ETDEWEB)

Barbis, D.P.; Maher, J.K.; Stanek, J.; Klaunberg, B.A.; Antczak, D.F.

1994-12-31

Two full-length clones encoding MHC class I genes were isolated by screening a horse cDNA library, using a probe encoding in human HLA-A2.2Y allele. The library was made in the pcDNA1 vector (Invitrogen, San Diego, CA), using mRNA from peripheral blood lymphocytes obtained from a Thoroughbred stallion (No. 0834) homozygous for a common horse MHC haplotype (ELA-A2, -B2, -D2; Antczak et al. 1984; Donaldson et al. 1988). The clones were sequenced, using SP6 and T7 universal primers and horse-specific oligonucleotides designed to extend previously determined sequences.

Molecular and Biological Characterization of an Isolate of Cucumber mosaic virus from Glycine soja by Generating its Infectious Full-genome cDNA Clones

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Mi Sa Vo Phan

2014-06-01

Full Text Available Molecular and biological characteristics of an isolate of Cucumber mosaic virus (CMV from Glycine soja (wild soybean, named as CMV-209, was examined in this study. Comparison of nucleotide sequences and phylogenetic analyses of CMV-209 with the other CMV strains revealed that CMV-209 belonged to CMV subgroup I. However, CMV-209 showed some genetic distance from the CMV strains assigned to subgroup IA or subgroup IB. Infectious full-genome cDNA clones of CMV-209 were generated under the control of the Cauliflower mosaic virus 35S promoter. Infectivity of the CMV-209 clones was evaluated in Nicotiana benthamiana and various legume species. Our assays revealed that CMV-209 could systemically infect Glycine soja (wild soybean and Pisum sativum (pea as well as N. benthamiana, but not the other legume species.

cDNA - ASTRA | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available ontents List of cDNA in locus Data file File name: astra_cdna.zip File URL: ftp://ftp.biosciencedbc.jp/archive/astra/LATEST/astra_cdn...a.zip File size: 3.3 MB Simple search URL http://togodb.biosciencedbc.jp/togodb/view/astra_cdna...n, Department of Molecular Genetics, National Institute of Agrobiological Sciences (Kikuchi et al., 2003; ftp://cdna

Semiochemicals from herbivory induced cotton plants enhance the foraging behavior of the cotton boll weevil, Anthonomus grandis.

Science.gov (United States)

Magalhães, D M; Borges, M; Laumann, R A; Sujii, E R; Mayon, P; Caulfield, J C; Midega, C A O; Khan, Z R; Pickett, J A; Birkett, M A; Blassioli-Moraes, M C

2012-12-01

The boll weevil, Anthonomus grandis, has been monitored through deployment of traps baited with aggregation pheromone components. However, field studies have shown that the number of insects caught in these traps is significantly reduced during cotton squaring, suggesting that volatiles produced by plants at this phenological stage may be involved in attraction. Here, we evaluated the chemical profile of volatile organic compounds (VOCs) emitted by undamaged or damaged cotton plants at different phenological stages, under different infestation conditions, and determined the attractiveness of these VOCs to adults of A. grandis. In addition, we investigated whether or not VOCs released by cotton plants enhanced the attractiveness of the aggregation pheromone emitted by male boll weevils. Behavioral responses of A. grandis to VOCs from conspecific-damaged, heterospecific-damaged (Spodoptera frugiperda and Euschistus heros) and undamaged cotton plants, at different phenological stages, were assessed in Y-tube olfactometers. The results showed that volatiles emitted from reproductive cotton plants damaged by conspecifics were attractive to adults boll weevils, whereas volatiles induced by heterospecific herbivores were not as attractive. Additionally, addition of boll weevil-induced volatiles from reproductive cotton plants to aggregation pheromone gave increased attraction, relative to the pheromone alone. The VOC profiles of undamaged and mechanically damaged cotton plants, in both phenological stages, were not different. Chemical analysis showed that cotton plants produced qualitatively similar volatile profiles regardless of damage type, but the quantities produced differed according to the plant's phenological stage and the herbivore species. Notably, vegetative cotton plants released higher amounts of VOCs compared to reproductive plants. At both stages, the highest rate of VOC release was observed in A. grandis-damaged plants. Results show that A. grandis uses

Sequential Sampling Plan of Anthonomus grandis (Coleoptera: Curculionidae) in Cotton Plants.

Science.gov (United States)

Grigolli, J F J; Souza, L A; Mota, T A; Fernandes, M G; Busoli, A C

2017-04-01

The boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), is one of the most important pests of cotton production worldwide. The objective of this work was to develop a sequential sampling plan for the boll weevil. The studies were conducted in Maracaju, MS, Brazil, in two seasons with cotton cultivar FM 993. A 10,000-m2 area of cotton was subdivided into 100 of 10- by 10-m plots, and five plants per plot were evaluated weekly, recording the number of squares with feeding + oviposition punctures of A. grandis in each plant. A sequential sampling plan by the maximum likelihood ratio test was developed, using a 10% threshold level of squares attacked. A 5% security level was adopted for the elaboration of the sequential sampling plan. The type I and type II error used was 0.05, recommended for studies with insects. The adjustment of the frequency distributions used were divided into two phases, so that the model that best fit to the data was the negative binomial distribution up to 85 DAE (Phase I), and from there the best fit was Poisson distribution (Phase II). The equations that define the decision-making for Phase I are S0 = -5.1743 + 0.5730N and S1 = 5.1743 + 0.5730N, and for the Phase II are S0 = -4.2479 + 0.5771N and S1 = 4.2479 + 0.5771N. The sequential sampling plan developed indicated the maximum number of sample units expected for decision-making is ∼39 and 31 samples for Phases I and II, respectively. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Assessment of wood density of seven clones of Eucalyptus grandis ...

African Journals Online (AJOL)

With the objective of evaluating the correlation of wood basic density with age in seven Eucalyptus grandis clones planted in Brazil, five trees in each clone were sampled at the ages of 0, 5, 1, 5, 2, 5, 3, 5, 4, 5 and 7, 5 years. The analysis of these samples showed that the intraclonal variation of the basic density (except for 0, ...

[Preparation of the cDNA microarray on the differential expressed cDNA of senescence-accelerated mouse's hippocampus].

Science.gov (United States)

Cheng, Xiao-Rui; Zhou, Wen-Xia; Zhang, Yong-Xiang

2006-05-01

Alzheimer' s disease (AD) is the most common form of dementia in the elderly. AD is an invariably fatal neurodegenerative disorder with no effective treatment. Senescence-accelerated mouse prone 8 (SAMP8) is a model for studying age-related cognitive impairments and also is a good model to study brain aging and one of mouse model of AD. The technique of cDNA microarray can monitor the expression levels of thousands of genes simultaneously and can be used to study AD with the character of multi-mechanism, multi-targets and multi-pathway. In order to disclose the mechanism of AD and find the drug targets of AD, cDNA microarray containing 3136 cDNAs amplified from the suppression subtracted cDNA library of hippocampus of SAMP8 and SAMR1 was prepared with 16 blocks and 14 x 14 pins, the housekeeping gene beta-actin and G3PDH as inner conference. The background of this microarray was low and unanimous, and dots divided evenly. The conditions of hybridization and washing were optimized during the hybridization of probe and target molecule. After the data of hybridization analysis, the differential expressed cDNAs were sequenced and analyzed by the bioinformatics, and some of genes were quantified by the real time RT-PCR and the reliability of this cDNA microarray were validated. This cDNA microarray may be the good means to select the differential expressed genes and disclose the molecular mechanism of SAMP8's brain aging and AD.

The oropharyngeal morphology in the semiaquatic giant Asian pond turtle, Heosemys grandis, and its evolutionary implications.

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Monika Lintner

Full Text Available The oropharynx as a functional entity plays a fundamental role in feeding. Transitions from aquatic to terrestrial lifestyles in vertebrates demanded major changes of the oropharynx for the required adaptations to a different feeding environment. Extant turtles evolved terrestrial feeding modes in three families (testudinids, emydids, geoemydids-independently from other amniotes-and are therefore important model organisms to reconstruct morpho-functional changes behind aquatic-terrestrial transitions. In this study we hypothesized that the oropharyngeal morphology in semiaquatic turtles of the geoemydid family shows parallels to testudinids, the only purely terrestrial extant lineage. We provide an in-depth description of the oropharynx in the semiaquatic geoemydid Heosemys grandis by using a combination of micro computed tomography (micro-CT and subsequent digital in situ 3-D reconstruction, scanning electron microscopy (SEM, and histology. We show that H. grandis has a large tongue with rough papillose surface and well-developed lingual muscles. The attachment sites of the lingual muscles on the hyolingual skeleton and their courses within the tongue are nearly identical with testudinids. The hyolingual skeleton itself is mainly cartilaginous and shows distinct-but compared to testudinids rather small-anterior extensions of the hyoid body and hypoglossum. Oral glands are well developed in H. grandis but are smaller and simpler than in testudinids. Similarly, oropharyngeal keratinization was minimal and found only in the anterior palate, regions close to the beak, and tongue tip. We conclude that H. grandis shows distinct oropharyngeal morpho-functional adaptations for a terrestrial lifestyle but still retains characters typical for aquatic forms. This makes this species an important example showing the oropharyngeal adaptations behind aquatic-terrestrial transitions in turtles.

Cloning of human tumor necrosis factor (TNF) receptor cDNA and expression of recombinant soluble TNF-binding protein

International Nuclear Information System (INIS)

Gray, P.W.; Barrett, K.; Chantry, D.; Turner, M.; Feldmann, M.

1990-01-01

The cDNA for one of the receptors for human tumor necrosis factor (TNF) has been isolated. This cDNA encodes a protein of 455 amino acids that is divided into an extracellular domain of 171 residues and a cytoplasmic domain of 221 residues. The extracellular domain has been engineered for expression in mammalian cells, and this recombinant derivative binds TNFα with high affinity and inhibits its cytotoxic activity in vitro. The TNF receptor exhibits similarity with a family of cell surface proteins that includes the nerve growth factor receptor, the human B-cell surface antigen CD40, and the rat T-cell surface antigen OX40. The TNF receptor contains four cysteine-rich subdomains in the extracellular portion. Mammalian cells transfected with the entire TNF receptor cDNA bind radiolabeled TNFα with an affinity of 2.5 x 10 -9 M. This binding can be competitively inhibited with unlabeled TNFα or lymphotoxin (TNFβ)

Cloning of Human Tumor Necrosis Factor (TNF) Receptor cDNA and Expression of Recombinant Soluble TNF-Binding Protein

Science.gov (United States)

Gray, Patrick W.; Barrett, Kathy; Chantry, David; Turner, Martin; Feldmann, Marc

1990-10-01

The cDNA for one of the receptors for human tumor necrosis factor (TNF) has been isolated. This cDNA encodes a protein of 455 amino acids that is divided into an extracellular domain of 171 residues and a cytoplasmic domain of 221 residues. The extracellular domain has been engineered for expression in mammalian cells, and this recombinant derivative binds TNFα with high affinity and inhibits its cytotoxic activity in vitro. The TNF receptor exhibits similarity with a family of cell surface proteins that includes the nerve growth factor receptor, the human B-cell surface antigen CD40, and the rat T-cell surface antigen OX40. The TNF receptor contains four cysteine-rich subdomains in the extra-cellular portion. Mammalian cells transfected with the entire TNF receptor cDNA bind radiolabeled TNFα with an affinity of 2.5 x 10-9 M. This binding can be competitively inhibited with unlabeled TNFα or lymphotoxin (TNFβ).

In vitro antioxidant activities of the fractions of Coccinia grandis l. leaf ...

African Journals Online (AJOL)

The present study was aimed at investigating the antioxidant activities of the various fractions of the hydromethanolic extract of the leaves of Coccinia grandis L. Voigt. (Cucurbitaceae). The antioxidant activities of the fractions have been evaluated by using nine in vitro assays and were compared to standard antioxidants ...

Procedure for normalization of cDNA libraries

Science.gov (United States)

Bonaldo, Maria DeFatima; Soares, Marcelo Bento

1997-01-01

This invention provides a method to normalize a cDNA library constructed in a vector capable of being converted to single-stranded circles and capable of producing complementary nucleic acid molecules to the single-stranded circles comprising: (a) converting the cDNA library in single-stranded circles; (b) generating complementary nucleic acid molecules to the single-stranded circles; (c) hybridizing the single-stranded circles converted in step (a) with complementary nucleic acid molecules of step (b) to produce partial duplexes to an appropriate Cot; (e) separating the unhybridized single-stranded circles from the hybridized single-stranded circles, thereby generating a normalized cDNA library.

Molecular cloning and characterization of a cDNA encoding the gibberellin biosynthetic enzyme ent-kaurene synthase B from pumpkin (Cucurbita maxima L.).

Science.gov (United States)

Yamaguchi, S; Saito, T; Abe, H; Yamane, H; Murofushi, N; Kamiya, Y

1996-08-01

The first committed step in the formation of diterpenoids leading to gibberellin (GA) biosynthesis is the conversion of geranylgeranyl diphosphate (GGDP) to ent-kaurene. ent-Kaurene synthase A (KSA) catalyzes the conversion of GGDP to copalyl diphosphate (CDP), which is subsequently converted to ent-kaurene by ent-kaurene synthase B (KSB). A full-length KSB cDNA was isolated from developing cotyledons in immature seeds of pumpkin (Cucurbita maxima L.). Degenerate oligonucleotide primers were designed from the amino acid sequences obtained from the purified protein to amplify a cDNA fragment, which was used for library screening. The isolated full-length cDNA was expressed in Escherichia coli as a fusion protein, which demonstrated the KSB activity to cyclize [3H]CDP to [3H]ent-kaurene. The KSB transcript was most abundant in growing tissues, but was detected in every organ in pumpkin seedlings. The deduced amino acid sequence shares significant homology with other terpene cyclases, including the conserved DDXXD motif, a putative divalent metal ion-diphosphate complex binding site. A putative transit peptide sequence that may target the translated product into the plastids is present in the N-terminal region.

Isolation and identification of the human homolog of a new p53-binding protein, Mdmx

NARCIS (Netherlands)

Shvarts, A.; Bazuine, M.; Dekker, P.; Ramos, Y. F.; Steegenga, W. T.; Merckx, G.; van Ham, R. C.; van der Houven van Oordt, W.; van der Eb, A. J.; Jochemsen, A. G.

1997-01-01

We recently reported the identification of a mouse cDNA encoding a new p53-associating protein that we called Mdmx because of its structural similarity to Mdm2, a well-known p53-binding protein. Here we report the isolation of a cDNA encoding the human homolog of Mdmx. The ORF of the cDNA encodes a

Divergence of host range and biological properties between natural isolate and full-length infectious cDNA clone of the Beet mild yellowing virus 2ITB.

Science.gov (United States)

Klein, Elodie; Brault, Véronique; Klein, Delphine; Weyens, Guy; Lefèbvre, Marc; Ziegler-Graff, Véronique; Gilmer, David

2014-01-01

Plant infection by poleroviruses is restricted to phloem tissues, preventing any classical leaf rub inoculation with viral RNA or virions. Efficient virus inoculation to plants is achieved by viruliferous aphids that acquire the virus by feeding on infected plants. The use of promoter-driven infectious cDNA is an alternative means to infect plants and allows reverse genetic studies to be performed. Using Beet mild yellowing virus isolate 2ITB (BMYV-2ITB), we produced a full-length infectious cDNA clone of the virus (named BMYV-EK) placed under the control of the T7 RNA polymerase and the Cauliflower mosaic virus 35S promoters. Infectivity of the engineered BMYV-EK virus was assayed in different plant species and compared with that of the original virus. We showed that in vitro- or in planta-derived transcripts were infectious in protoplasts and in whole plants. Importantly, the natural aphid vector Myzus persicae efficiently transmitted the viral progeny produced in infected plants. By comparing agroinoculation and aphid infection in a host range assay, we showed that the engineered BMYV-EK virus displayed a similar host range to BMYV-2ITB, except for Nicotiana benthamiana, which proved to be resistant to systemic infection with BMYV-EK. Finally, both the BMYV-EK P0 and the full-length clone were able to strongly interfere with post-transcriptional gene silencing. © 2013 BSPP AND JOHN WILEY & SONS LTD.

cDNA cloning and immunological characterization of the rye grass allergen Lol p I.

Science.gov (United States)

Perez, M; Ishioka, G Y; Walker, L E; Chesnut, R W

1990-09-25

The complete amino acid sequence of two "isoallergenic" forms of Lol p I, the major rye grass (Lolium perenne) pollen allergen, was deduced from cDNA sequence analysis. cDNA clones isolated from a Lolium perenne pollen library contained an open reading frame coding for a 240-amino acid protein. Comparison of the nucleotide and deduced amino acid sequence of two of these clones revealed four changes at the amino acid level and numerous nucleotide differences. Both clones contained one possible asparagine-linked glycosylation site. Northern blot analysis shows one RNA species of 1.2 kilobases. Based on the complete amino acid sequence of Lol p I, overlapping peptides covering the entire molecule were synthesized. Utilizing these peptides we have identified a determinant within the Lol p I molecule that is recognized by human leukocyte antigen class II-restricted T cells obtained from persons allergic to rye grass pollen.

Isolation and preliminary function analysis of a Na + /H + antiporter ...

African Journals Online (AJOL)

A full-length cDNA Na+/H+ antiporter gene (MzNHX1) was isolated from Malus zumi according to the homologous Na+/H+ antiporter gene region in plants. Sequence analysis indicated that the cDNA was 2062 bp in length, including an open reading frame (ORF) of 1629 bp, which encoded a predicted polypeptide of 542 ...

(TECTONA GRANDIS LEAF POWDER

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Yash Mishra

2015-01-01

Full Text Available In this study, the adsorption potential of Teak (Tectona grandis leaf powder (TLP toremove Methylene blue (MB and Malachite Green (MG dye molecules from aqueoussolution was investigated. Batch experiments were conducted to evaluate the influenceof operational parameters such as, pH (2−9, adsorbent dosage (1−7 g/L, contact time(15−150 minutes and initial dye concentration (20−120 mg/L at stirring speed of 150rpm for the adsorption of MB and MG on TLP. Maximum removal efficiency of 98.4%and 95.1% was achieved for MB and MG dye, respectively. The experimentalequilibrium data were analysed using Langmuir, Freundlich and Temkin isothermmodels and it was found that, it fitted well to the Freundlich isotherm model. Thesurface structure and morphology of the adsorbent was characterized using scanningelectron microscopy (SEM and the presence of functional groups and its interactionwith the dye molecules were analysed using Fourier transform infrared spectroscopy(FTIR. Based on the investigation, it has been demonstrated that the teak leaf powderhas good potential for effective adsorption of methylene blue and malachite green dye.

Lectin cDNA and transgenic plants derived therefrom

Science.gov (United States)

Raikhel, Natasha V.

2000-10-03

Transgenic plants containing cDNA encoding Gramineae lectin are described. The plants preferably contain cDNA coding for barley lectin and store the lectin in the leaves. The transgenic plants, particularly the leaves exhibit insecticidal and fungicidal properties.

Insulin-Mimetic Action of Rhoifolin and Cosmosiin Isolated from Citrus grandis (L. Osbeck Leaves: Enhanced Adiponectin Secretion and Insulin Receptor Phosphorylation in 3T3-L1 Cells

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Yerra Koteswara Rao

2011-01-01

Full Text Available Citrus grandis (L. Osbeck (red wendun leaves have been used in traditional Chinese medicine to treat several illnesses including diabetes. However, there is no scientific evidence supporting these actions and its active compounds. Two flavone glycosides, rhoifolin and cosmosiin were isolated for the first time from red wendun leaves and, identified these leaves are rich source for rhoifolin (1.1%, w/w. In differentiated 3T3-L1 adipocytes, rhoifolin and cosmosiin showed dose-dependent response in concentration range of o.oo1–5 μM and 1–20 μM, respectively, in biological studies beneficial to diabetes. Particularly, rhoifolin and cosmosiin at 0.5 and 20 μM, respectively showed nearly similar response to that 10 nM of insulin, on adiponectin secretion level. Furthermore, 5 μM of rhoifolin and 20 μM of cosmosiin showed equal potential with 10 nM of insulin to increase the phosphorylation of insulin receptor-β, in addition to their positive effect on GLUT4 translocation. These findings indicate that rhoifolin and cosmosiin from red wendun leaves may be beneficial for diabetic complications through their enhanced adiponectin secretion, tyrosine phosphorylation of insulin receptor-β and GLUT4 translocation.

cDNA table - RPD | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available of data contents Results of homology search to cDNA clones in the KOME. Data file File name: rpd_cdna.zip F...ile URL: ftp://ftp.biosciencedbc.jp/archive/rpd/LATEST/rpd_cdna.zip File size: 15 KB Simple search URL http:...//togodb.biosciencedbc.jp/togodb/view/rpd_cdna#en Data acquisition method - Data

Molecular cloning of cDNA for rat brain metallothionein-2 and regulation of its gene expression

Energy Technology Data Exchange (ETDEWEB)

Saijoh, Kiyofumi; Sumino, Kimiaki [Department of Public Health, Kobe University School of Medicine (Japan); Kuno, Takayoshi; Shuntoh, Hisato; Tanaka, Chikako [Department of Pharmacology, Kobe University of Medicine (Japan)

1989-01-01

A rat brain metallothionein-II (MT-II) complementary DNA (cDNA) clone was isolated from a cDNA plasmid library, which was prepared from non-treated rat brain mRNA, by a colony screening procedure using /sup 32/P-labeled synthetic oligonucleotide probes. It is deduced that the clone encodes for a protein of 61 amino acids comprising 20 cysteines, which is highly homologous to MT-IIs in other species. Northern blot analysis demonstrated major mRNA species in the brain, liver and kidneys (approximately 350 b in size), which is induced in response to dexamethasone, zinc, cadmium and mercury but not to methyl mercury. These findings confirm that MT-II genes are expressed and regulated both by steroid and heavy metals in the brain as well as in peripheral organs. (author).

Molecular cloning of cDNA for rat brain metallothionein-2 and regulation of its gene expression

International Nuclear Information System (INIS)

Saijoh, Kiyofumi; Sumino, Kimiaki; Kuno, Takayoshi; Shuntoh, Hisato; Tanaka, Chikako

1989-01-01

A rat brain metallothionein-II (MT-II) complementary DNA (cDNA) clone was isolated from a cDNA plasmid library, which was prepared from non-treated rat brain mRNA, by a colony screening procedure using 32 P-labeled synthetic oligonucleotide probes. It is deduced that the clone encodes for a protein of 61 amino acids comprising 20 cysteines, which is highly homologous to MT-IIs in other species. Northern blot analysis demonstrated major mRNA species in the brain, liver and kidneys (approximately 350 b in size), which is induced in response to dexamethasone, zinc, cadmium and mercury but not to methyl mercury. These findings confirm that MT-II genes are expressed and regulated both by steroid and heavy metals in the brain as well as in peripheral organs. (author)

Dynamic transcriptome profiling of the floral buds in the dioecious cucurbit Coccinia grandis using RNA-Seq

Directory of Open Access Journals (Sweden)

Jatindra Nath Mohanty

2017-10-01

Full Text Available Angiosperms exhibits diversified sexual systems encompassing bisexual, monoecious and dioecious conditions. Dioecy offers opportunities to explore separately, the male and female systems giving an insight into the evolutionary, developmental and molecular processes of sex expression in plants. Coccinia grandis (Family: Cucurbitaceae with small genome size and heteromorphic sex chromosomes is often considered a model dioecious plant for sex evolution. However, the information relating to its genetic orientation, physical state and sex determining factors is highly ambiguous and limited. In the present study we have attempted to identify the molecular basis of sex determination in C. grandis through genome wide transcriptome profiling of the floral buds. About 75 million clean reads were generated resulting in 72,479 unigenes for male library and 63,308 unigenes for female library with a mean length of 736 bp. 1410 unigenes were differentially expressed (DEGs between the male and female buds as identified from the RNA-Seq pattern and qRT-PCR validation. Functional annotation using BLAST2GO and KEGG revealed high enrichment of DEGs in phytohormone biosynthesis, hormone signaling and transduction, transcriptional regulation and methyl transferase activity. Manifold up-regulation of genes phytohormone responsive genes such as ARF6, ACC synthase1, SNRK2 and BRI1-associated receptor kinase 1 (BAK1 suggest that a signaling crosstalk is implicated in the sex determination of this species. Besides, a wide range of transcription factors including zinc fingers, homeodomain leucine zippers and MYBs were recognized as major determinants of male specific expression in the dioecious plant. Additionally, C. grandis transcriptome revealed 48 target genes for many miRNAs sequences with established role in floral development and sex determination. Overall, our study resulted in the identification of a large amount of molecular resources that could be critical to

PARASITISM OF BOLL WEEVIL (Anthonomus grandis IN FLOWER BUDS OF COTTON PLANT, IN THE MUNICIPAL DISTRICT OF GOIÂNIA-GO PARASITISMO DO BICUDO DO ALGODOEIRO (Anthonomus grandis EM BOTÕES FLORAIS DO ALGODOEIRO, NO MUNICÍPIO DE GOIÂNIA-GO

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Paulo Marçal Fernandes

2007-09-01

Full Text Available

This work studied the indexes of parasitism of A. grandis in floral buttons of the cotton plants, collected in the soil and in the plants, in an area not treat with insecticides, located in the School of Agronomy of the Universidade Federal de Goiás, municipal district of Goiânia-GO. Floral buttons were collected with and without sign of oviparousness of the beaked ones. They presented larger parasitism occurrence in those collected in the soil. The parasites were identified as: Chelonus sp. (Microchelonus, Bracon sp. and Pteromalidae.

KEY-WORDS: Insecta; parasitism; cotton plant; Anthonomus grandis.

Estudou-se o índice de parasitismo de A. grandis em botões florais de algodoeiro coletados no solo e nas plantas, em uma área não tratada com inseticidas, localizada na Escola de Agronomia da Universidade Federal de Goiás, no município de Goiânia (GO. Foram coletados botões florais com e sem puncturas de oviposição dos bicudos. Verificou-se um maior parasitismo nos botões florais coletados no solo. Os parasitóides foram identificados como Chelonus sp. (Microchelonus, Bracon sp. e Pteromalidae.

PALAVRAS-CHAVE: Insecta; parasitismo; algodoeiro; Anthonomus grandis.

Performance of Australian provenances of Eucalyptus grandis and Eucalyptus saligna in Hawaii

Science.gov (United States)

Roger G. Skolmen

1986-01-01

Australian provenances of Eucalyptus grandis and E. saligna were compared at four locations on the island of Hawaii to seek seed sources better than those in current use which were introduced earlier from unrecorded locations in Australia. A broad range of latitude and elevation was represented among the provenances. At all four...

Toxicity to cotton boll weevil Anthonomus grandis of a trypsin inhibitor from chickpea seeds.

Science.gov (United States)

de P G Gomes, Angélica; Dias, Simoni C; Bloch, Carlos; Melo, Francislete R; Furtado, José R; Monnerat, Rose G; Grossi-de-Sá, Maria F; Franco, Octávio L

2005-02-01

Cotton (Gossypium hirsutum L.) is an important agricultural commodity, which is attacked by several pests such as the cotton boll weevil Anthonomus grandis. Adult A. grandis feed on fruits and leaf petioles, reducing drastically the crop production. The predominance of boll weevil digestive serine proteinases has motivated inhibitor screenings in order to discover new ones with the capability to reduce the digestion process. The present study describes a novel proteinase inhibitor from chickpea seeds (Cicer arietinum L.) and its effects against A. grandis. This inhibitor, named CaTI, was purified by using affinity Red-Sepharose Cl-6B chromatography, followed by reversed-phase HPLC (Vydac C18-TP). SDS-PAGE and MALDI-TOF analyses, showed a unique monomeric protein with a mass of 12,877 Da. Purified CaTI showed significant inhibitory activity against larval cotton boll weevil serine proteinases (78%) and against bovine pancreatic trypsin (73%), when analyzed by fluorimetric assays. Although the molecular mass of CaTI corresponded to alpha-amylase/trypsin bifunctional inhibitors masses, no inhibitory activity against insect and mammalian alpha-amylases was observed. In order to observe CaTI in vivo effects, an inhibitor rich fraction was added to an artificial diet at different concentrations. At 1.5% (w/w), CaTI caused severe development delay, several deformities and a mortality rate of approximately 45%. These results suggested that CaTI could be useful in the production of transgenic cotton plants with enhanced resistance toward cotton boll weevil.

Antraquinonas e naftoquinonas do caule de um espécime de reflorestamento de Tectona grandi (Verbenaceae

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Rafael Y.O. Moreira

Full Text Available O fracionamento do extrato hexânico do caule de um espécime de reflorestamento de Tectona grandis (Verbenaceae, através de procedimentos fitoquímicos clássicos, levou ao isolamento das naftoquinonas lapachol e desidro-a-lapachona e das antraquinonas tectoquinona e obtusifolina. As estruturas das substâncias foram caracterizadas através da análise de métodos espectrométricos de RMN. Este é o primeiro estudo fitoquímico de um espécime de reflorestamento de Tectona grandis, no Brasil, sendo o objetivo principal deste trabalho a comprovação da presença de tectoquinona em espécimes cultivados.

Preparation of fluorescent-dye-labeled cDNA from RNA for microarray hybridization.

Science.gov (United States)

Ares, Manuel

2014-01-01

This protocol describes how to prepare fluorescently labeled cDNA for hybridization to microarrays. It consists of two steps: first, a mixture of anchored oligo(dT) and random hexamers is used to prime amine-modified cDNA synthesis by reverse transcriptase using a modified deoxynucleotide with a reactive amine group (aminoallyl-dUTP) and an RNA sample as a template. Second, the cDNA is purified and exchanged into bicarbonate buffer so that the amine groups in the cDNA react with the dye N-hydroxysuccinimide (NHS) esters, covalently joining the dye to the cDNA. The dye-coupled cDNA is purified again, and the amount of dye incorporated per microgram of cDNA is determined.

Constructing and detecting a cDNA library for mites.

Science.gov (United States)

Hu, Li; Zhao, YaE; Cheng, Juan; Yang, YuanJun; Li, Chen; Lu, ZhaoHui

2015-10-01

RNA extraction and construction of complementary DNA (cDNA) library for mites have been quite challenging due to difficulties in acquiring tiny living mites and breaking their hard chitin. The present study is to explore a better method to construct cDNA library for mites that will lay the foundation on transcriptome and molecular pathogenesis research. We selected Psoroptes cuniculi as an experimental subject and took the following steps to construct and verify cDNA library. First, we combined liquid nitrogen grinding with TRIzol for total RNA extraction. Then, switching mechanism at 5' end of the RNA transcript (SMART) technique was used to construct full-length cDNA library. To evaluate the quality of cDNA library, the library titer and recombination rate were calculated. The reliability of cDNA library was detected by sequencing and analyzing positive clones and genes amplified by specific primers. The results showed that the RNA concentration was 836 ng/μl and the absorbance ratio at 260/280 nm was 1.82. The library titer was 5.31 × 10(5) plaque-forming unit (PFU)/ml and the recombination rate was 98.21%, indicating that the library was of good quality. In the 33 expressed sequence tags (ESTs) of P. cuniculi, two clones of 1656 and 1658 bp were almost identical with only three variable sites detected, which had an identity of 99.63% with that of Psoroptes ovis, indicating that the cDNA library was reliable. Further detection by specific primers demonstrated that the 553-bp Pso c II gene sequences of P. cuniculi had an identity of 98.56% with those of P. ovis, confirming that the cDNA library was not only reliable but also feasible.

Molecular cloning of growth hormone encoding cDNA of Indian

Indian Academy of Sciences (India)

A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences of Labeo rohita, Cirrhina mrigala and Catla catla have been cloned, characterized and overexpressed in ...

Cloning, characterization and heterologous expression of epoxide hydrolase-encoding cDNA sequences from yeasts belonging to the genera Rhodotorula and Rhodosporidium

NARCIS (Netherlands)

Visser, H.; Weijers, C.A.G.M.; Ooyen, van A.J.J.; Verdoes, J.C.

2002-01-01

Epoxide hydrolase-encoding cDNA sequences were isolated from the basidiomycetous yeast species Rhodosporidium toruloides CBS 349, Rhodosporidium toruloides CBS 14 and Rhodotorula araucariae CBS 6031 in order to evaluate the molecular data and potential application of this type of enzymes. The

[Cloning of cDNA for RNA polymerase subunit from the fission yeast Schizosaccharomyces pombe by heterospecific complementation in Saccharomyces cerevisiae].

Science.gov (United States)

Shpakovskiĭ, G V; Lebedenko, E N; Thuriaux, P

1997-02-01

The rpb10 cDNA of the fission yeast Schizosaccharomyces pombe, encoding one of the five small subunits common to all three nuclear DNA-dependent RNA polymerases, was isolated from an expression cDNA library by two independent approaches: PCR-based screening and direct suppression by means of heterospecific complementation of a temperature-sensitive mutant defective in the corresponding gene of Saccharomyces cerevisiae. The cloned Sz. pombe cDNA encodes a protein Rpb10 of 71 amino acids with an M of 8,275 Da, sharing 51 amino acids (71% identity) with the subunit ABC10 beta of RNA polymerases I-III from S. cerevisiae. All eukaryotic members of this protein family have the same general organization featuring two highly conserved motifs (RCFT/SCGK and RYCCRRM) around an atypical zinc finger and an additional invariant HVDLIEK motif toward the C-terminal end. The last motif is only characteristics for homologs from eukaryotes. In keeping with this remarkable structural conservation, the Sz. pombe cDNA also fully complemented a S. cerevisiae deletion mutant lacking subunit ABC10 beta (null allele rpb10-delta 1::HIS3).

Cloning and analysis of the mouse Fanconi anemia group a cDNA and an overlapping penta zinc finger cDNA

NARCIS (Netherlands)

Wong, JCY; Alon, N; Norga, K; Kruyt, FAE; Youssoufian, H; Buchwald, M

2000-01-01

Despite the cloning of four disease-associated genes for Fanconi anemia (FA), the molecular pathogenesis of FA remains largely unknown. To study FA complementation group A using the mouse as a mode I system, we cloned and characterized the mouse homolog of the human FANCA cDNA, The mouse cDNA

Utilização de madeiras de Eucalyptus grandis E Eucalyptus dunnii para produção de painéis de partículas orientadas – OSB

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Setsuo Iwakiri

2008-01-01

Full Text Available This study was developed to evaluate the feasibility of OSB manufacturing using woods of Eucalyptus grandis and Eucalyptus dunnii. Boards with nominal density of 0,70 g/cm³ and 1,0 g/cm³ were manufactured in laboratory, using 100% of wood particles from Pinus taeda, Eucalyptus grandis and Eucalyptus dunnii, and mixtures of 50% of Pinus taeda in the internal layer of the board, with 50% of Eucalyptus grandis and 50% of Eucalyptus dunnii. The boards of Eucalyptus grandis with density of 0,70 g/cm³, as standard board density, showed the values of properties compatible with the requirements of the Canadian and European Standards and also in relation of boards manufactured from Pinus taeda. The results of the mechanical properties showed an increase in the MOE and MOR in static bending with the increase in the board density, opening the possibility to use the high density OSB for applications requiring higher strength. The results of this research indicate that wood of Eucalyptus grandis can be used as alternative specie to OSB manufacturing in the Brazil.

cDNA library construction of two human Demodexspecies.

Science.gov (United States)

Niu, DongLing; Wang, RuiLing; Zhao, YaE; Yang, Rui; Hu, Li; Lei, YuYang; Dan, WeiChao

2017-06-01

The research of Demodex, a type of pathogen causing various dermatoses in animals and human beings, is lacking at RNA level. This study aims at extracting RNA and constructing cDNA library for Demodex. First, P. cuniculiand D. farinaewere mixed to establish homogenization method for RNA extraction. Second, D. folliculorumand D. breviswere collected and preserved in Trizol, which were mixed with D. farinaerespectively to extract RNA. Finally, cDNA library was constructed and its quality was assessed. The results indicated that for D. folliculorum& D. farinae, the recombination rate of cDNA library was 90.67% and the library titer was 7.50 × 104 pfu/ml. 17 of the 59 positive clones were predicted to be of D. folliculorum; For D. brevis& D. farinae, the recombination rate was 90.96% and the library titer was 7.85 x104 pfu/ml. 40 of the 59 positive clones were predicted to be of D. brevis. Further detection by specific primers demonstrated that mtDNA cox1, cox3and ATP6 detected from cDNA libraries had 96.52%-99.73% identities with the corresponding sequences in GenBank. In conclusion, the cDNA libraries constructed for Demodexmixed with D. farinaewere successful and could satisfy the requirements for functional genes detection.

Isolation of a novel abscisic acid stress ripening ( OsASR ) gene ...

African Journals Online (AJOL)

Isolation of a novel abscisic acid stress ripening ( OsASR ) gene from rice and analysis of the response of this gene to abiotic stresses. ... The cDNA with the whole open reading frame (ORF) was amplified by PCR and cloned. Sequence analysis showed that the cDNA encodes a protein of 284 amino acid residues with ...

Tectona grandis L. f (the teak tree; Hindi: Sagallll or Segllll) (~r ...

Indian Academy of Sciences (India)

Tectona grandis L. f (the teak tree; Hindi: Sagallll or Segllll) (~r Verhell({Ceae is a large deciduous tree cllitivatedfor its timber. Leaves are large; flmvers are small, white, svveet-scented and borne on highly branched inflorescences. Fruit is hard and enveloped by bladder-like cal.vx. The timber is one of the best for all wood ...

Genetic and environmental factors affecting rooting in Eucalyptus Grandis X Eucalyptus Longistrata hybrid cuttings

CSIR Research Space (South Africa)

Naidoo, N

2012-03-01

Full Text Available An investigation was undertaken to study the rooting ability of E. grandis x E. longirostrata hybrid cuttings. The plant material was sourced from five families in seedling derived hedges at two nurseries, as well as five families coppiced from...

cDNA library information - Dicty_cDB | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available List Contact us Dicty_cDB cDNA library information Data detail Data name cDNA library information DOI 10.189...s Data item Description cDNA library name Names of cDNA libraries (AF, AH, CF, CH, FC, FC-IC, FCL, SF, SH, S...(C) 5) sexually fusion-competent KAX3 cells (Gamete phase) (F) cDNA library construction method How to construct cDNA library...dir) 2) Full-length cDNA libraries (oligocapped method)(fl) 3) Gamete-specific subtraction library (sub) cDNA library... construction protocol Link to the webpage describing the protocol for generating cDNA library Size

Avaliação do teor de umidade da madeira de Eucalyptus grandis por medidores elétricos resistivos. Evaluation of the Eucalyptus grandis lumber moisture content by resistancetype moisture meters.

Directory of Open Access Journals (Sweden)

João Eduardo Guarnetti dos SANTOS

2006-01-01

Full Text Available O presente estudo teve como objetivoverificar a precisão de dois tipos de medidoreselétricos de teor de umidade durante o processo desecagem da madeira de Eucalyptus grandis. Foramretiradas amostras representativas de 14 tábuas deEucalyptus grandis e secas em estufa elétrica a40 ºC de temperatura, até que o material atingisse10% de umidade. Durante a secagem foramdeterminados, periodicamente, o teor de umidadeatravés do método de massas correntes e deverificações simultâneas com um medidor elétricoportátil (EMM e com o sistema de controle de umsecador convencional (KCS. Os resultados mostraramque: (1 o sensor de umidade KCS pode substituiro método gravimétrico durante a secagem damadeira; (2 o medidor do teor de umidade EMMsubestima os reais teores de umidade durante asecagem da madeira e não é indicado parasubstituir o método gravimétrico de determinação de umidade.The aim of the study was to evaluate theprecision of two types of electric moisture metersduring the drying process of Eucalyptus grandisboards. Samples were obtained from 14 boards ofEucalyptus grandis and they were dried in electriclaboratory oven at 40 ºC of temperature, until thewood achieve 10% of moisture content. During thedrying, the moisture content was determined bygravimetric method and simultaneous checks by anelectric moisture meter (EMM and by kilncontrol system (KCS. The results showed that:(1 the KCS can replace the gravimetric methodduring the wood drying; (2 the EMMunderestimate the real moisture content during thedrying of boards and it is not indicate as substituteof the gravimetric method.

cDNA library Table - KAIKOcDNA | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available c00951-005 Description of data contents List of Bombyx mori cDNA libraries. Data file File name: kaiko_cdna_...library.zip File URL: ftp://ftp.biosciencedbc.jp/archive/kaiko-cdna/LATEST/kaiko_cdna_library.zip File size:... 4.8 KB Simple search URL http://togodb.biosciencedbc.jp/togodb/view/kaiko_cdna_l

Binding proteins for the regulatory subunit (RII-B) of brain cAMP-dependent protein kinase II: isolation and initial characterization of cDNA clones

International Nuclear Information System (INIS)

Bregman, D.B.; Hu, E.; Rubin, C.S.

1987-01-01

In mammalian brain several proteins bind RII-B with high affinity. An example is P75, which co-purifies with RII-B and also complexes Ca 2+ -calmodulin. Thus, RII-B binding proteins (RBPs) might play a role in integrating the Ca 2+ and cAMP signalling pathways in the CNS. In order to study the structure and function of these polypeptides they have isolated cloned cDNAs for RBPs by screening brain λgt11 expression libraries using a functional assay: the binding of 32 P-labeled RII to fusion proteins produced by recombinants expressing RII binding domains. Inserts from rat brain recombinant clones λ7B and λ10B both hybridize to a brain mRNA of 7000 nucleotides. Northern gel analyses indicate that the putative RBP mRNA is also expressed in lung, but not in several other tissues. The λ7B insert was subcloned into the expression plasmid pINIA. A 50 kDa high affinity RII-B binding polypeptide accumulated in E. coli transformed with pINIA-7B. Two RBP cDNAs (λ77, λ100A) have been retrieved from a bovine λgt 11 library using a monoclonal antibody directed against P75 and the binding assay respectively. On Southern blots the insert from λ100A hybridizes to the cDNA insert from clones λ77, suggesting that λ 77 cDNA might contain sequences coding for both an RII binding domain and a P75 epitope. The bovine λ100A insert also hybridizes with the rat λ7B clone indicating that an RII binding domain is conserved in the two species

Método de diagnóstico para el monitoreo de resistencia a insecticidas en poblaciones de "picudo del algodonero", Anthonomus grandis (Coleoptera: Curculionidae A diagnostic test for insecticide resistance monitoring in "cotton boll weevil" Anthonomus grandis (Coleoptera: Curculionidae populations

Directory of Open Access Journals (Sweden)

Teodoro Stadler

2009-06-01

Full Text Available El control de las poblaciones de Anthonomus grandis Boheman, por debajo de su umbral de daño económico durante el ciclo del cultivo del algodón, se realiza en forma efectiva hasta el momento, a través de insecticidas de síntesis. La presión selectiva de las aplicaciones extensivas e intensivas de insecticidas hace imperativa la detección temprana de focos de resistencia a los mismos, en función de un correcto manejo del fenómeno. Se desarrolló un método de diagnóstico de resistencia para A. grandis a partir de la técnica "vial test", que fue adaptada en forma de "kit" para el monitoreo rápido y sencillo de los focos de resistencia en el campo. La toxicidad (CL99, para calcular la concentración discriminante (CD del insecticida y la preparación del "kit", se obtiene a partir de bioensayos de laboratorio con una cepa normal susceptible de A. grandis. Se determinó la vida media de los insecticidas dentro de los viales por CIPAC MT 46, para establecer una fecha de vencimiento del "kit". La CD y el método en su conjunto fueron validados a través de ensayos a campo. El "kit", usado en el monitoreo de resistencia en el campo, fue especialmente diseñado para ser utilizado en las condiciones geográficas, económicas y socio-culturales presentes en la región algodonera argentina. La implementación de esta técnica permitirá conseguir la información necesaria, y así obtener una apropiada alternancia de insecticidas. Como consecuencia, se prevé una reducción de impacto ambiental de las prácticas agronómicas en el control de plagas en algodón.The in-season control of the cotton boll weevil Anthonomus grandis Boheman is done by insecticide application, which so far is the only effective way to reduce boll weevil populations to levels below economic significance. The extensive and intensive control actions with insecticides cause selective pressure on pest populations. Thus, to achieve an accurate insecticide resistance

Isolation, cDNA cloning and gene expression of an antibacterial protein from larvae of the coconut rhinoceros beetle, Oryctes rhinoceros.

Science.gov (United States)

Yang, J; Yamamoto, M; Ishibashi, J; Taniai, K; Yamakawa, M

1998-08-01

An antibacterial protein, designated rhinocerosin, was purified to homogeneity from larvae of the coconut rhinoceros beetle, Oryctes rhinoceros immunized with Escherichia coli. Based on the amino acid sequence of the N-terminal region, a degenerate primer was synthesized and reverse-transcriptase PCR was performed to clone rhinocerosin cDNA. As a result, a 279-bp fragment was obtained. The complete nucleotide sequence was determined by sequencing the extended rhinocerosin cDNA clone by 5' rapid amplification of cDNA ends. The deduced amino acid sequence of the mature portion of rhinocerosin was composed of 72 amino acids without cystein residues and was shown to be rich in glycine (11.1%) and proline (11.1%) residues. Comparison of the deduced amino acid sequence of rhinocerosin with those of other antibacterial proteins indicated that it has 77.8% and 44.6% identity with holotricin 2 and coleoptrecin, respectively. Rhinocerosin had strong antibacterial activity against E. coli, Streptococcus pyogenes, Staphylococcus aureus but not against Pseudomonas aeruginosa. Results of reverse-transcriptase PCR analysis of gene expression in different tissues indicated that the rhinocerosin gene is strongly expressed in the fat body and the Malpighian tubule, and weakly expressed in hemocytes and midgut. In addition, gene expression was inducible by bacteria in the fat body, the Malpighian tubule and hemocyte but constitutive expression was observed in the midgut.

Cloning of the cDNA and gene for a human D2 dopamine receptor

International Nuclear Information System (INIS)

Grady, D.K.; Makam, H.; Stofko, R.E.; Bunzow, J.R.; Civelli, O.; Marchionni, M.A.; Alfano, M.; Frothingham, L.; Fischer, J.B.; Burke-Howie, K.J.; Server, A.C.

1989-01-01

A clone encoding a human D 2 dopamine receptor was isolated from a pituitary cDNA library and sequenced. The deduced protein sequence is 96% identical with that of the cloned rat receptor with one major difference: the human receptor contains an additional 29 amino acids in its putative third cytoplasmic loop. Southern blotting demonstrated the presence of only one human D 2 receptor gene. Two overlapping phage containing the gene were isolated and characterized. DNA sequence analysis of these clones showed that the coding sequence is interrupted by six introns and that the additional amino acids present in the human pituitary receptor are encoded by a single exon of 87 base pairs. The involvement of this sequence in alternative splicing and its biological significance are discussed

Temporal dynamics of the response to Al stress in Eucalyptus grandis × Eucalyptus camaldulensis

Directory of Open Access Journals (Sweden)

Berenice K. de Alcântara

2015-06-01

Full Text Available Lipid peroxidation and root elongation of Eucalyptus grandis × Eucalyptus camaldulensis were studied under stress conditions in response to aluminum (Al, a metal known to limit agricultural productivity in acidic soils primarily due to reduced root elongation. In Brazil, the Grancam 1277 hybrid (E. grandis × E. camaldulensis has been planted in the "Cerrado", a region of the country with a wide occurrence of acidic soils. The present study demonstrated that the hybrid exhibited root growth reduction and increased levels of lipid peroxidation after 24h of treatment with 100 µM of Al, which was followed by a reduction in lipid peroxidation levels and the recovery of root elongation after 48h of Al exposure, suggesting a rapid response to the early stressful conditions induced by Al. The understanding of the temporal dynamics of Al tolerance may be useful for selecting more tolerant genotypes and for identifying genes of interest for applications in bioengineering.

Molecular cloning of a catalase cDNA from Nicotiana glutinosa L. and its repression by tobacco mosaic virus infection.

Science.gov (United States)

Yi, S Y; Yu, S H; Choi, D

1999-06-30

Recent reports revealed that catalase has a role in the plant defense mechanism against a broad range of pathogens through being inhibited by salicylic acid (SA). During an effort to clone disease resistance-responsive genes, a cDNA encoding catalase (Ngcat1; Nicotiana glutinosa cat1) was isolated from a tobacco cDNA library. In N. glutinosa, catalase is encoded by a small gene family. The deduced amino acid sequence of the Ngcat1 cDNA has 98% homology with the cat1 gene of N. plumbaginifolia. The Ngcat1 expression is controlled by the circadian clock, and its mRNA level is the most abundant in leaves. Both the expression of Ngcat1 mRNA and its enzyme activity in the tobacco plant undergoing a hypersensitive response (HR) to TMV infection were repressed. The repression of the mRNA level was also observed following treatment with SA. These results imply that SA may act as an inhibitor of catalase transcription during the HR of tobacco. Cloning and expression of the Ngcat1 in tobacco following pathogen infection and SA treatment are presented.

Characterization of the porcine carboxypeptidase E cDNA

DEFF Research Database (Denmark)

Hreidarsdôttir, G.E.; Cirera, Susanna; Fredholm, Merete

2007-01-01

the sequence of the cDNA for the porcine CPE gene including all the coding region and the 3'-UTR region was generated. Comparisons with bovine, human, mouse, and rat CPE cDNA sequences showed that the coding regions of the gene are highly conserved both at the nucleotide and at the amino acid level. A very low...

Fiscal 2000 report on result of the full-length cDNA structure analysis; 2000 nendo kanzen cho cDNA kozo kaiseki seika hokokusho

Energy Technology Data Exchange (ETDEWEB)

NONE

2001-03-01

This paper explains the results of research on full-length cDNA structure analysis for the period from April, 2000 to March, 2001. The outline of human genome sequence was published in June, 2000. In Japan, human gene analysis was such that, as the basic technology of the bio industry, a millennium project was decided in the budget of fiscal 2000. The full-length cDNA structure analysis is the core of the project. The libraries of cDNA were prepared using full-length and more than 4-5kbp-long cDNAs by oligo-capping method. It began from determining partial sequence data at end cDNA, and then, with new clones selected therefrom, full-length human cDNA sequence data were determined. The partial sequence data determined by fiscal 2000 were 1,035,000 clones while the full-length sequence data were 12,144 clones. The sequence data obtained were analyzed by homology search and translated into amino acid coding sequences, with predictions conducted on protein functions. A clustering method was examined that selects new clones from partial sequences. Database was constructed on gene expression profiles and disease-related gene sequence data. (NEDO)

Characterization of a pollen-specific cDNA clone from Nicotiana tabacum expressed during microgametogenesis and germination.

Science.gov (United States)

Weterings, K; Reijnen, W; van Aarssen, R; Kortstee, A; Spijkers, J; van Herpen, M; Schrauwen, J; Wullems, G

1992-04-01

This report describes the isolation and characterization of a cDNA clone representing a gene specifically expressed in pollen. A cDNA library was constructed against mRNA from mature pollen of Nicotiana tabacum. It was screened differentially against cDNA from mRNA of leaf and of pollen. One clone, NTPc303, was further characterized. On northern blot this clone hybridizes to a transcript 2100 nucleotides in length. NTPc303 is abundant in pollen. Expression of the corresponding gene is restricted to pollen, because no other generative or vegetative tissue contains transcripts hybridizing to NTPc303. Expression of NTP303 is evolutionarily conserved: homologous transcripts are present in pollen from various plant species. The first NTP303 transcripts are detectable on northern blot at the early bi-nucleate stage and accumulate until the pollen has reached maturity. During germination and pollen tube growth in vitro new NTP303 transcripts appear. This transcription has been proved by northern blots as well as by pulse labelling experiments. Nucleotide sequence analysis revealed that NTPc303 has an open reading frame coding for a predicted protein of 62 kDa. This protein shares homology to ascorbate oxidase and other members of the blue copper oxidase family. A possible function for this clone during pollen germination is discussed.

Cloning of the γ-aminobutyric acid (GABA) ρ1 cDNA: A GABA receptor subunit highly expressed in the retina

International Nuclear Information System (INIS)

Cutting, G.R.; Lu, Luo; Kasch, L.M.; Montrose-Rafizadeh, C.; Antonarakis, S.E.; Guggino, W.B.; Kazazian, H.H. Jr.; O'Hara, B.F.; Donovan, D.M.; Shimada, Shoichi; Uhl, G.R.

1991-01-01

Type A γ-aminobutyric acid (GABA A ) receptors are a family of ligand-gated chloride channels that are the major inhibitory neurotransmitter receptors in the nervous system. Molecular cloning has revealed diversity in the subunits that compose this heterooligomeric receptor, but each previously elucidated subunit displays amino acid similarity in conserved structural elements. The authors have used these highly conserved regions to identify additional members of this family by using the polymerase chain reaction (PCR). One PCR product was used to isolate a full-length cDNA from a human retina cDNA library. The mature protein predicted from this cDNA sequence is 458 amino acids long and displays between 30 and 38% amino acid similarity to the previously identified GABA A subunits. This gene is expressed primarily in the retina but transcripts are also detected in the brain, lung, and thymus. Injection of Xenopus oocytes with RNA transcribed in vitro produces a GABA-responsive chloride conductance and expression of the cDNA in COS cells yields GABA-displaceable muscimol binding. These features are consistent with our identification of a GABA subunit, GABA ρ 1 , with prominent retinal expression that increases the diversity and tissue specificity of this ligand-gated ion-channel receptor family

Human liver phosphatase 2A: cDNA and amino acid sequence of two catalytic subunit isotypes

International Nuclear Information System (INIS)

Arino, J.; Woon, Chee Wai; Brautigan, D.L.; Miller, T.B. Jr.; Johnson, G.L.

1988-01-01

Two cDNA clones were isolated from a human liver library that encode two phosphatase 2A catalytic subunits. The two cDNAs differed in eight amino acids (97% identity) with three nonconservative substitutions. All of the amino acid substitutions were clustered in the amino-terminal domain of the protein. Amino acid sequence of one human liver clone (HL-14) was identical to the rabbit skeletal muscle phosphatase 2A cDNA (with 97% nucleotide identity). The second human liver clone (HL-1) is encoded by a separate gene, and RNA gel blot analysis indicates that both mRNAs are expressed similarly in several human clonal cell lines. Sequence comparison with phosphatase 1 and 2A indicates highly divergent amino acid sequences at the amino and carboxyl termini of the proteins and identifies six highly conserved regions between the two proteins that are predicted to be important for phosphatase enzymatic activity

Midgut GPI-anchored proteins with alkaline phosphatase activity from the cotton boll weevil (Anthonomus grandis) are putative receptors for the Cry1B protein of Bacillus thuringiensis.

Science.gov (United States)

Martins, Erica Soares; Monnerat, Rose Gomes; Queiroz, Paulo Roberto; Dumas, Vinicius Fiuza; Braz, Shélida Vasconcelos; de Souza Aguiar, Raimundo Wagner; Gomes, Ana Cristina Menezes Mendes; Sánchez, Jorge; Bravo, Alejandra; Ribeiro, Bergmann Morais

2010-02-01

Cry toxins from Bacillus thuringiensis (Bt) are used for insect control. They interact with specific receptors located on the host cell surface and are activated by host proteases following receptor binding resulting in midgut epithelial cells lysis. In this work we had cloned, sequenced and expressed a cry1Ba toxin gene from the B thuringiensis S601 strain which was previously shown to be toxic to Anthonomus grandis, a cotton pest. The Cry1Ba6 protein expressed in an acrystaliferous B. thuringiensis strain was toxic to A. grandis in bioassays. The binding of Cry1Ba6 toxin to proteins located in the midgut brush border membrane of A. grandis was analyzed and we found that Cry1Ba6 binds to two proteins (62 and 65kDa) that showed alkaline phosphatase (ALP) activity. This work is the first report that shows the localization of Cry toxin receptors in the midgut cells of A. grandis. 2009. Published by Elsevier Ltd.

Efeito da hidrólise ácida dos taninos de Eucalyptus grandis w. hill ex Maiden nas propriedades dos adesivos tânicos Effect of the acid hydrolyses of Eucalyptus grandis w. hill ex Maiden tannins in the properties of the tannic adhesives

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Angélica de Cássia Oliveira Carneiro

2009-08-01

Full Text Available Este trabalho teve como objetivo avaliar o efeito da hidrólise ácida sobre as propriedades e resistência ao cisalhamento dos adesivos de taninos, extraídos a partir da casca de Eucalyptus grandis W. Hill ex Maiden. Os adesivos foram sintetizados com taninos hidrolisados, empregando-se quatro valores de pH, três tempos de reação e 10% de formaldeído em relação à massa seca de taninos. Foram produzidas 96 juntas coladas, constituídas de duas lâminas de madeira de Eucalyptus grandis. As resistências ao cisalhamento e falha na madeira foram determinadas de acordo com a norma ASTM D 2339-93. Concluiu-se que a hidrólise ácida dos taninos reduziu a viscosidade dos adesivos e aumentou a resistência ao cisalhamento na linha de cola. De modo geral, as juntas coladas apresentaram baixo percentual de falha na madeira.The objective of this research was to evaluate the effect of the acid hydrolysis of Eucalyptus grandis W. Hill ex Maiden bark tannins on the properties of tannic adhesives. Adhesives were synthesized with tannins hydrolyzed at four pH values, three reaction times and 10% of formaldehyde based on the tannin dry weight. Ninety-six glued joints were prepared with Eucalyptus grandis thin boards. Shear resistance and wood failure percentage were determined according to the ASTM D 2339-93 standards. It was concluded that tannin hydrolysis decreased adhesive viscosity and increased the glue line shear resistance. However, it was observed a low percentage of wood failure.

A Polymerase Chain Reaction-Based Method for Isolating Clones from a Complimentary DNA Library in Sheep

Science.gov (United States)

Friis, Thor Einar; Stephenson, Sally; Xiao, Yin; Whitehead, Jon

2014-01-01

The sheep (Ovis aries) is favored by many musculoskeletal tissue engineering groups as a large animal model because of its docile temperament and ease of husbandry. The size and weight of sheep are comparable to humans, which allows for the use of implants and fixation devices used in human clinical practice. The construction of a complimentary DNA (cDNA) library can capture the expression of genes in both a tissue- and time-specific manner. cDNA libraries have been a consistent source of gene discovery ever since the technology became commonplace more than three decades ago. Here, we describe the construction of a cDNA library using cells derived from sheep bones based on the pBluescript cDNA kit. Thirty clones were picked at random and sequenced. This led to the identification of a novel gene, C12orf29, which our initial experiments indicate is involved in skeletal biology. We also describe a polymerase chain reaction-based cDNA clone isolation method that allows the isolation of genes of interest from a cDNA library pool. The techniques outlined here can be applied in-house by smaller tissue engineering groups to generate tools for biomolecular research for large preclinical animal studies and highlights the power of standard cDNA library protocols to uncover novel genes. PMID:24447069

Lufenuron impact upon Anthonomus grandis Boheman (Coleoptera: Curculionidae) midgut and its reflection in gametogenesis.

Science.gov (United States)

Costa, Hilton Nobre; da Cunha, Franklin Magliano; Cruz, Glaucilane Santos; D'assunção, Carolline Guimarães; Rolim, Guilherme Gomes; Barros, Maria Edna Gomes; Breda, Mariana Oliveira; Teixeira, Alvaro Aguiar Coelho; Teixeira, Valéria Wanderley

2017-04-01

The insecticide Match® (lufenuron), one of the main insect growth regulators used in pest control, has been presented as a viable alternative against the boll weevil, Anthonomus grandis (Coleoptera: Curculionidae), by inhibiting chitin synthesis. Thus, this study aimed to examine whether Match® interferes in the synthesis of the peritrophic matrix, leading to changes in the midgut epithelium, resulting in nutritional deficiency and reflecting, thereby, in the gametogenesis process of A. grandis. Floral cotton buds were immersed in the insecticide solution (800μL of Match®+200mL of distilled water) and offered to the adult insects. The midguts of the insects were evaluated after 24 and 120h after feeding. The gonads were evaluated after 120h. The results showed that Match®, in both evaluation periods, induced histopathological alterations such as disorganization, vacuolization and desquamation of the midgut epithelium; histochemical modifications in the distribution patterns of carbohydrates, although without quantitative changes; and a strong decrease in protein levels. No apoptosis were observed, however, there was an increase in the number of regenerative cell nests. In the testicles, a reduction in the amount of spermatozoids and reduced carbohydrate levels were observed, but no difference in protein levels. The ovarioles presented structural disorganization of follicular cells, yolk reduction and decrease in protein levels, however, no change in carbohydrates levels was noted. Therefore, it is concluded that Match® performs histopathologic and histochemical alterations in the midgut epithelium and the gonads of A. grandis adults, reflecting in the gametogenesis process, presenting itself as a promising tool in the management of this pest on cotton crops. Copyright © 2016 Elsevier Inc. All rights reserved.

Nucleases as a barrier to gene silencing in the cotton boll weevil, Anthonomus grandis.

Science.gov (United States)

Almeida Garcia, Rayssa; Lima Pepino Macedo, Leonardo; Cabral do Nascimento, Danila; Gillet, François-Xavier; Moreira-Pinto, Clidia Eduarda; Faheem, Muhammad; Moreschi Basso, Angelina Maria; Mattar Silva, Maria Cristina; Grossi-de-Sa, Maria Fatima

2017-01-01

RNA interference (RNAi) approaches have been applied as a biotechnological tool for controlling plant insect pests via selective gene down regulation. However, the inefficiency of RNAi mechanism in insects is associated with several barriers, including dsRNA delivery and uptake by the cell, dsRNA interaction with the cellular membrane receptor and dsRNA exposure to insect gut nucleases during feeding. The cotton boll weevil (Anthonomus grandis) is a coleopteran in which RNAi-mediated gene silencing does not function efficiently through dsRNA feeding, and the factors involved in the mechanism remain unknown. Herein, we identified three nucleases in the cotton boll weevil transcriptome denoted AgraNuc1, AgraNuc2, and AgraNuc3, and the influences of these nucleases on the gene silencing of A. grandis chitin synthase II (AgraChSII) were evaluated through oral dsRNA feeding trials. A phylogenetic analysis showed that all three nucleases share high similarity with the DNA/RNA non-specific endonuclease family of other insects. These nucleases were found to be mainly expressed in the posterior midgut region of the insect. Two days after nuclease RNAi-mediated gene silencing, dsRNA degradation by the gut juice was substantially reduced. Notably, after nucleases gene silencing, the orally delivered dsRNA against the AgraChSII gene resulted in improved gene silencing efficiency when compared to the control (non-silenced nucleases). The data presented here demonstrates that A. grandis midgut nucleases are effectively one of the main barriers to dsRNA delivery and emphasize the need to develop novel RNAi delivery strategies focusing on protecting the dsRNA from gut nucleases and enhancing its oral delivery and uptake to crop insect pests.

MPO cDNA clone identifies an RFLP with PstI

Energy Technology Data Exchange (ETDEWEB)

Miki, T; Weil, S C; Rosner, G L; Reid, M S; Kidd, K K

1988-02-25

A myeloperoxidase (MPO) cDNA clone (pHMP7: 270 base pair insert in the vector pGEM-1reverse arrow was isolated from a library created from human promyelocytic (HL-60) cell mRNA. PstI (CTGCA/G) (New England Biolabs) identifies a simple two-allele polymorphism with bands at either 2.2 kb (Al) or 2.0 kb (A2). There are three constant bands at 2.8 kb, 0.95 kb and 0.6 kb. Preliminary family data show evidence of linkage to several markers in proximal 17q, with MPO closest to the Growth Hormone cluster at 17q22-q24. Autosomal condominant segregation was observed in four large reference pedigrees with several informative matings.

Aplicação de uma técnica alternativa de manejo físico do solo no cultivo de Eucalyptus grandis W.Hill (Myrtaceae Application of an alternative technique for physical soil management in cultivation of Eucalyptus grandis W.Hill (Myrtaceae

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João Paulo de Maçaneiro

2013-02-01

Full Text Available O objetivo deste estudo foi avaliar o processo de crescimento do Eucalyptus grandis quando submetido à irregularização do terreno. Baseando-se na "Ótica da Teoria do Caos" e partindo-se da hipótese de que as áreas reflorestadas por E. grandis são consideravelmente sensíveis às condições iniciais de preparação do solo, aplicou-se a técnica das rugosidades (variações do relevo alternando superfícies côncavas e convexas para desencadear ao longo do tempo propriedades emergentes que aceleram o processo de crescimento vegetal. A área de estudo localiza-se na Bacia Hidrográfica do Rio Itajaí, em Brusque, SC. Esta foi dividida em quatro parcelas menores: duas com tratamentos irregulares (IR-A e IR-B e outras duas com tratamentos regulares (R-A e R-B. Os tratamentos irregulares consistiram na abertura de cavas, utilizando-se uma retroescavadeira hidráulica, intercaladas com 1 m de largura, 4 a 5 m de comprimento e 0,5 m de profundidade. Nos tratamentos regulares foi adotado o cultivo mínimo do solo, onde o preparo do solo ficou restrito às linhas ou covas de plantio. Na análise do desenvolvimento de E. grandis (altura, diâmetro do colo e na altura do peito - DAP verificou-se diferenças estatísticas entre as técnicas de preparação do solo, sendo os maiores valores nos tratamentos irregulares. Nas parcelas irregulares (IR-A e IR-B foram encontrados os maiores valores médios de altura (5,29 m e 5,46 m, diâmetro do colo (45,65 mm e 45,4 mm e DAP (4,44 cm e 4,79 cm, respectivamente. Pressupõe-se que as rugosidades funcionaram efetivamente como componentes auxiliares na internalização da matéria, retendo água, sedimentos e nutrientes, fato que deve ter potencializado e acelerado o crescimento do E. grandis.The aim of this study was to evaluate the process of growth of Eucalyptus grandis non-regularization when subjected to the ground. Relying on "Optical Chaos Theory" and starting from the assumption that the reforested

Antioxidant, antiglycation and insulinotrophic properties of Coccinia grandis (L. in vitro: Possible role in prevention of diabetic complications

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Packirisamy Meenatchi

2017-01-01

Full Text Available In an attempt to develop Complementary and Alternative Medicine (CAM for the treatment of diabetes and related complications, the antidiabetic potential of the mature unripe fruits of Coccinia grandis (CGF was evaluated. Oxidative stress and glycation plays an important role in manifesting of diabetes and vascular complications. Agents with antioxidant and antiglycation properties may retard these pathological alterations. In this study, the edible plant Coccinia grandis was assessed for in vitro estimation of antioxidant and antiglycation potential and its insulinotrophic properties in RINm5F cells. Antioxidant activity was evaluated as DPPH (1,1-diphenyl-2-picrylhydrazyl, hydrogen peroxide and superoxide anion scavenging activities, whereas the protein glycation inhibitory potential was evaluated using in vitro albumin-fructose glycation model. Glycation inhibition was estimated by different biochemical parameters viz. fructosamine, protein carbonyl group and protein aggregation using thioflavin T fluorescence. C. grandis extract exerted a dose dependent radical scavenging activity and exhibited a significant antiglycation potential. The extract also showed a significant insulinotrophic property with 1.28 and 1.71-fold increase in insulin release when compared to control at 0.25 and 0.50 mg/mL, respectively. These data suggest the possible antidiabetic role of CGF extract, presumably by its antioxidant, antiglycation and insulin secretory effects. Present findings provide experimental evidence that the fruits of C. grandis have potential antidiabetic activity which might be used as a functional food and safe remedy for the treatment of diabetes and associated complications. This study also revealed that the plant can be a promising source for development of natural antiglycating agents and novel insulin secretagogues.

Cloning of the cDNA for U1 small nuclear ribonucleoprotein particle 70K protein from Arabidopsis thaliana

Science.gov (United States)

Reddy, A. S.; Czernik, A. J.; An, G.; Poovaiah, B. W.

1992-01-01

We cloned and sequenced a plant cDNA that encodes U1 small nuclear ribonucleoprotein (snRNP) 70K protein. The plant U1 snRNP 70K protein cDNA is not full length and lacks the coding region for 68 amino acids in the amino-terminal region as compared to human U1 snRNP 70K protein. Comparison of the deduced amino acid sequence of the plant U1 snRNP 70K protein with the amino acid sequence of animal and yeast U1 snRNP 70K protein showed a high degree of homology. The plant U1 snRNP 70K protein is more closely related to the human counter part than to the yeast 70K protein. The carboxy-terminal half is less well conserved but, like the vertebrate 70K proteins, is rich in charged amino acids. Northern analysis with the RNA isolated from different parts of the plant indicates that the snRNP 70K gene is expressed in all of the parts tested. Southern blotting of genomic DNA using the cDNA indicates that the U1 snRNP 70K protein is coded by a single gene.

Identification of PEG-induced water stress responsive transcripts using co-expression network in Eucalyptus grandis.

Science.gov (United States)

Ghosh Dasgupta, Modhumita; Dharanishanthi, Veeramuthu

2017-09-05

Ecophysiological studies in Eucalyptus have shown that water is the principal factor limiting stem growth. Effect of water deficit conditions on physiological and biochemical parameters has been extensively reported in Eucalyptus. The present study was conducted to identify major polyethylene glycol induced water stress responsive transcripts in Eucalyptus grandis using gene co-expression network. A customized array representing 3359 water stress responsive genes was designed to document their expression in leaves of E. grandis cuttings subjected to -0.225MPa of PEG treatment. The differentially expressed transcripts were documented and significantly co-expressed transcripts were used for construction of network. The co-expression network was constructed with 915 nodes and 3454 edges with degree ranging from 2 to 45. Ninety four GO categories and 117 functional pathways were identified in the network. MCODE analysis generated 27 modules and module 6 with 479 nodes and 1005 edges was identified as the biologically relevant network. The major water responsive transcripts represented in the module included dehydrin, osmotin, LEA protein, expansin, arabinogalactans, heat shock proteins, major facilitator proteins, ARM repeat proteins, raffinose synthase, tonoplast intrinsic protein and transcription factors like DREB2A, ARF9, AGL24, UNE12, WLIM1 and MYB66, MYB70, MYB 55, MYB 16 and MYB 103. The coordinated analysis of gene expression patterns and coexpression networks developed in this study identified an array of transcripts that may regulate PEG induced water stress responses in E. grandis. Copyright © 2017 Elsevier B.V. All rights reserved.

Green synthesis of silver nanoparticles mediated by Coccinia grandis and Phyllanthus emblica: a comparative comprehension

Science.gov (United States)

Nayagam, Vasanth; Gabriel, Melchias; Palanisamy, Kumaravel

2018-04-01

Fruit extracts also have the potentiality to synthesize silver nanoparticles, which serve as antimicrobial agent in the biological field. At present, the field of biomedical largely depends on the biosynthesized NPs to fight against the multi-drug-resistant pathogens. The fruit residue of Coccinia grandis and Phyllanthus emblica are employed for synthesizing AgNPs by green method. The NPs are further subjected to UV, FTIR, SEM, and XRD measurements. The ten different pathogens were tested against the AgNPs synthesized. The same were tested for early growth of some seed variety too, so as to check the advantages of AgNPs. The UV spectrum analysis showed 442 nm and 423 nm, respectively, and FTIR peaks for the functional group that is responsible for the conversion of NPs were observed at 1640.02 for N-H bond amines (Coccinia grandis) and at 1637.45 for N-H bond amines (Phyllanthus emblica). The SEM results also illustrated that AgNPs are spherical in shape. The XRD patterns indicate the crystalline nature of the AgNPs formed with both these plants. The antimicrobial assay of AgNPs from Coccinia grandis shows maximum zone of inhibition (14 mm) for Vibrio cholerae whereas the AgNPs from Phyllanthus emblica show maximum inhibition at distinct points, namely for Staphylococcus aureus, Vibrio cholerae, Salmonella typhi, and Proteus mirabilis (12 mm). Seed germination initiated by AgNPs is quiet effective and healthier compared to the water-induced seeds. Hence, biogenic AgNPs have various applications in favour of human society.

cDNA microarray screening in food safety

International Nuclear Information System (INIS)

Roy, Sashwati; Sen, Chandan K.

2006-01-01

The cDNA microarray technology and related bioinformatics tools presents a wide range of novel application opportunities. The technology may be productively applied to address food safety. In this mini-review article, we present an update highlighting the late breaking discoveries that demonstrate the vitality of cDNA microarray technology as a tool to analyze food safety with reference to microbial pathogens and genetically modified foods. In order to bring the microarray technology to mainstream food safety, it is important to develop robust user-friendly tools that may be applied in a field setting. In addition, there needs to be a standardized process for regulatory agencies to interpret and act upon microarray-based data. The cDNA microarray approach is an emergent technology in diagnostics. Its values lie in being able to provide complimentary molecular insight when employed in addition to traditional tests for food safety, as part of a more comprehensive battery of tests

cDNA structure, genomic organization and expression patterns of ...

African Journals Online (AJOL)

Visfatin was a newly identified adipocytokine, which was involved in various physiologic and pathologic processes of organisms. The cDNA structure, genomic organization and expression patterns of silver Prussian carp visfatin were described in this report. The silver Prussian carp visfatin cDNA cloned from the liver was ...

Mouse tetranectin: cDNA sequence, tissue-specific expression, and chromosomal mapping

DEFF Research Database (Denmark)

Ibaraki, K; Kozak, C A; Wewer, U M

1995-01-01

regulation, mouse tetranectin cDNA was cloned from a 16-day-old mouse embryo library. Sequence analysis revealed a 992-bp cDNA with an open reading frame of 606 bp, which is identical in length to the human tetranectin cDNA. The deduced amino acid sequence showed high homology to the human cDNA with 76......(s) of tetranectin. The sequence analysis revealed a difference in both sequence and size of the noncoding regions between mouse and human cDNAs. Northern analysis of the various tissues from mouse, rat, and cow showed the major transcript(s) to be approximately 1 kb, which is similar in size to that observed...

Sequencing and characterization of asclepain f: the first cysteine peptidase cDNA cloned and expressed from Asclepias fruticosa latex.

Science.gov (United States)

Trejo, Sebastián A; López, Laura M I; Caffini, Néstor O; Natalucci, Claudia L; Canals, Francesc; Avilés, Francesc X

2009-07-01

Asclepain f is a papain-like protease previously isolated and characterized from latex of Asclepias fruticosa. This enzyme is a member of the C1 family of cysteine proteases that are synthesized as preproenzymes. The enzyme belongs to the alpha + beta class of proteins, with two disulfide bridges (Cys22-Cys63 and Cys56-Cys95) in the alpha domain, and another one (Cys150-Cys201) in the beta domain, as was determined by molecular modeling. A full-length 1,152 bp cDNA was cloned by RT-RACE-PCR from latex mRNA. The sequence was predicted as an open reading frame of 340 amino acid residues, of which 16 residues belong to the signal peptide, 113 to the propeptide and 211 to the mature enzyme. The full-length cDNA was ligated to pPICZalpha vector and expressed in Pichia pastoris. Recombinant asclepain f showed endopeptidase activity on pGlu-Phe-Leu-p-nitroanilide and was identified by PMF-MALDI-TOF MS. Asclepain f is the first peptidase cloned and expressed from mRNA isolated from plant latex, confirming the presence of the preprocysteine peptidase in the latex.

A rapid method for screening arrayed plasmid cDNA library by PCR

International Nuclear Information System (INIS)

Hu Yingchun; Zhang Kaitai; Wu Dechang; Li Gang; Xiang Xiaoqiong

1999-01-01

Objective: To develop a PCR-based method for rapid and effective screening of arrayed plasmid cDNA library. Methods: The plasmid cDNA library was arrayed and screened by PCR with a particular set of primers. Results: Four positive clones were obtained through about one week. Conclusion: This method can be applied to screening not only normal cDNA clones, but also cDNA clones-containing small size fragments. This method offers significant advantages over traditional screening method in terms of sensitivity, specificity and efficiency

Determination of chemical elements in Eucalyptus grandis, manured with Ballad's, by neutrons activation analysis

International Nuclear Information System (INIS)

Mateus, Natalina de Fatima; Madi Filho, Tufic

2007-01-01

The biosolid is a mud resulting from the biological treatment of wasted liquids. It is considered as a profitable alternative and important to minimize the environmental impact generated by the sewage thrown in to sanitary lands, in forest cultures like the Eucalyptus grandis. The objective of this work was to detect which chemical elements are present in Eucalyptus grandis samples, fertilized with different quantities of biosolid. The eucalyptuses of Estacao Experimental de Ciencias Florestais of Itatinga were planted in March of 1998 and collected with five years old. The used biosolid was produced by Station of Treatment of Sewer of Barueri - SP, classified as kind B. For the determination of the presence and quantity of chemical elements in the eucalyptus samples, an analysis technique by neutronic activation (NAA) was used followed by gamma rays spectroscopy. The samples were irradiated in the Nuclear Reactor IEA-R1 of IPEN-SP, followed by the measure of induced gamma rays activity, using a Detector HPGe. The presence, mainly of Br, Mn, Na and K, was detected in all analyzed samples. (author)

Primary structure of bovine pituitary secretory protein I (chromogranin A) deduced from the cDNA sequence

International Nuclear Information System (INIS)

Ahn, T.G.; Cohn, D.V.; Gorr, S.U.; Ornstein, D.L.; Kashdan, M.A.; Levine, M.A.

1987-01-01

Secretory protein I (SP-I), also referred to as chromogranin A, is an acidic glycoprotein that has been found in every tissue of endocrine and neuroendocrine origin examined but never in exocrine or epithelial cells. Its co-storage and co-secretion with peptide hormones and neurotransmitters suggest that it has an important endocrine or secretory function. The authors have isolated cDNA clones from a bovine pituitary λgt11 expression library using an antiserum to parathyroid SP-I. The largest clone (SP4B) hybridized to a transcript of 2.1 kilobases in RNA from parathyroid, pituitary, and adrenal medulla. Immunoblots of bacterial lysates derived from SP4B lysognes demonstrated specific antibody binding to an SP4B/β-galactosidase fusion protein (160 kDa) with a cDNA-derived component of 46 kDa. Radioimmunoassay of the bacterial lystates with SP-I antiserum yielded parallel displacement curves of 125 I-labeled SP-I by the SP4B lysate and authentic SP-I. SP4B contains a cDNA of 1614 nucleotides that encodes a 449-amino acid protein (calculated mass, 50 kDa). The nucleotide sequences of the pituitary SP-I cDNA and adrenal medullary SP-I cDNAs are nearly identical. Analysis of genomic DNA suggests that pituitary, adrenal, and parathyroid SP-I are products of the same gene

Primary structure of bovine pituitary secretory protein I (chromogranin A) deduced from the cDNA sequence

Energy Technology Data Exchange (ETDEWEB)

Ahn, T.G.; Cohn, D.V.; Gorr, S.U.; Ornstein, D.L.; Kashdan, M.A.; Levine, M.A.

1987-07-01

Secretory protein I (SP-I), also referred to as chromogranin A, is an acidic glycoprotein that has been found in every tissue of endocrine and neuroendocrine origin examined but never in exocrine or epithelial cells. Its co-storage and co-secretion with peptide hormones and neurotransmitters suggest that it has an important endocrine or secretory function. The authors have isolated cDNA clones from a bovine pituitary lambdagt11 expression library using an antiserum to parathyroid SP-I. The largest clone (SP4B) hybridized to a transcript of 2.1 kilobases in RNA from parathyroid, pituitary, and adrenal medulla. Immunoblots of bacterial lysates derived from SP4B lysognes demonstrated specific antibody binding to an SP4B/..beta..-galactosidase fusion protein (160 kDa) with a cDNA-derived component of 46 kDa. Radioimmunoassay of the bacterial lystates with SP-I antiserum yielded parallel displacement curves of /sup 125/I-labeled SP-I by the SP4B lysate and authentic SP-I. SP4B contains a cDNA of 1614 nucleotides that encodes a 449-amino acid protein (calculated mass, 50 kDa). The nucleotide sequences of the pituitary SP-I cDNA and adrenal medullary SP-I cDNAs are nearly identical. Analysis of genomic DNA suggests that pituitary, adrenal, and parathyroid SP-I are products of the same gene.

Effects of soybean Kunitz trypsin inhibitor on the cotton boll weevil (Anthonomus grandis).

Science.gov (United States)

Franco, Octávio L; Dias, Simoni C; Magalhães, Claudio P; Monteiro, Ana C S; Bloch, Carlos; Melo, Francislete R; Oliveira-Neto, Osmundo B; Monnerat, Rose G; Grossi-de-Sá, Maria Fátima

2004-01-01

The cotton boll weevil, Anthonomus grandis, is an economically important pest of cotton in tropical and subtropical areas of several countries in the Americas, causing severe losses due to their damage in cotton floral buds. Enzymatic assays using gut extracts from larval and adult boll weevil have demonstrated the presence of digestive serine proteinase-like activities. Furthermore, in vitro assays showed that soybean Kunitz trypsin inhibitor (SKTI) was able to inhibit these enzymes. Previously, in vivo effects of black-eyed pea trypsin chymotrypsin inhibitor (BTCI) have been demonstrated towards the boll weevil pest. Here, when neonate larvae were reared on an artificial diet containing SKTI at three different concentrations, a reduction of larval weight of up to 64% was observed for highest SKTI concentration 500 microM. The presence of SKTI caused an increase in mortality and severe deformities of larvae, pupae and adult insects. This work therefore represents the first observation of a Kunitz trypsin inhibitor active in vivo and in vitro against A. grandis. Bioassays suggested that SKTI could be used as a tool in engineering crop plants, which might exhibit increased resistance against cotton boll weevil.

Magnesium alleviates adverse effects of lead on growth, photosynthesis and ultrastructural alterations of Torreya grandis seedlings

Directory of Open Access Journals (Sweden)

Jie Shen

2016-11-01

Full Text Available Magnesium (Mg2+ has been shown to reduce the physiological and biochemical stress in plants caused by heavy metals. To date our understanding of how Mg2+ ameliorates the adverse effects of heavy metals in plants is scarce. The potential effect of Mg2+ on lead (Pb2+ toxicity in plants has not yet been studied. This study was designed to clarify the mechanism of Mg2+-induced alleviation of lead (Pb2+ toxicity. Torreya grandis (T. grandis seedlings were grown in substrate contaminated with 0, 700 and 1400 mg Pb2+ per kg-1 and with or without the addition of 1040 mg kg-1 Mg2+. Growth parameters, concentrations of Pb2+ and Mg2+ in the plants’ shoots and roots, photosynthetic pigment, gas exchange parameters, the maximum quantum efficiency (Fv/Fm, root oxidative activity, ultrastructure of chloroplasts and root growth were determined to analyze the effect of different Pb2+ concentrations in the seedlings as well as the potential ameliorating effect of Mg2+ on the Pb2+ induced toxicity. The growth of T. grandis seedlings cultivated in soils treated with 1400 mg kg-1 Pb2+ was significantly reduced compared with that of plants cultivated in soils treated with 0 or 700 mg kg-1 Pb2+. The addition of 1040 mg kg-1 Mg2+ improved the growth of the Pb2+-stressed seedlings, which was accompanied by increased chlorophyll content, the net photosynthetic rate and Fv/Fm, and enhanced chloroplasts development. In addition, the application of Mg2+ induced plants to accumulate five times higher concentrations of Pb2+ in the roots and to absorb and translocate four times higher concentrations of Mg2+ to the shoots than those without Mg2+ application. Furthermore, Mg2+ addition increased root growth and oxidative activity, and protected the root ultrastructure. To the best of our knowledge, our study is the first report on the mechanism of Mg2+-induced alleviation of Pb2+ toxicity. The gener¬ated results may have important implications for understanding the

KARAKTERISTIK SEKUEN cDNA PENGKODE GEN ANTI VIRUS DARI UDANG WINDU, Penaeus monodon

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Andi Parenrengi

2016-11-01

Full Text Available Transgenesis pada ikan merupakan sebuah teknik modern yang berpotensi besar dalam menghasilkan organisme yang memiliki karakter lebih baik melalui rekombinan DNA gen target termasuk gen anti virus dalam peningkatan resistensi pada udang. Gen anti virus PmAV (Penaeus monodon Anti Viral gene merupakan salah satu gen pengkode anti virus yang berasal dari spesies krustase. Penelitian ini dilakukan untuk mengetahui karakteristik gen anti virus yang diisolasi dari udang windu, Penaeus monodon. Isolasi gen anti virus menggunakan metode Polymerase Chain Reaction (PCR dan selanjutnya dipurifikasi untuk sekuensing. Data yang dihasilkan dianalisis dengan program Genetyx Versi 7 dan basic local alignment search tool (BLAST. Hasil penelitian menunjukkan bahwa gen anti virus PmAV yang berhasil diisolasi dari cDNA udang windu dengan panjang sekuen 520 bp yang mengkodekan 170 asam amino. BLAST-N menunjukkan tingkat similaritas yang sangat tinggi (100% dengan gen anti virus yang ada di GeneBank. Komposisi asam amino penyusun gen anti virus yang paling besar adalah serin (10,00%, sedangkan yang terkecil adalah asam amino prolin dan lisin masing-masing 1,76%. Analisis sekuen gen dan deduksi asam amino (BLAST-P memperlihatkan adanya C-type lectin-like domain (CTLD yang memiliki kemiripan dengan gen C-type lectin yang diisolasi dari beberapa spesies krustase. Transgenic fish technology is a potential modern technique in producing better character organism through DNA recombinant of target genes including anti viral gene for improvement of shrimp immunity. PmAV (Penaeus monodon Anti Viral gene is one of anti viral genes isolated from crustacean species. The research was conducted to analyze the characteristics anti viral gene isolated from tiger prawn, Penaeus monodon. Anti viral gene was isolated using Polymerase Chain Reaction (PCR technique and then purified for sequencing. Data obtained were analyzed using Genetyx Version 7 software and basic local alignment

Complementation of radiation-sensitive Ataxia telangiectasia cells after transfection of cDNA expression libraries and cosmid clones from wildtype cells

International Nuclear Information System (INIS)

Fritz, E.

1994-06-01

In this Ph.D.-thesis, phenotypic complementation of AT-cells (AT5BIVA) by transfection of cDNA-expression-libraries was adressed: After stable transfection of cDNA-expression-libraries G418 resistant clones were selected for enhanced radioresistance by a fractionated X-ray selection. One surviving transfectant clone (clone 514) exhibited enhanced radiation resistance in dose-response experiments and further X-ray selections. Cell cycle analysis revealed complementation of untreated and irradiated 514-cells in cell cycle progression. The rate of DNA synthesis, however, is not diminished after irradiation but shows the reverse effect. A transfected cDNA-fragment (AT500-cDNA) was isolated from the genomic DNA of 514-cells and proved to be an unknown DNA sequence. A homologous sequence could be detected in genomic DNA from human cell lines, but not in DNA from other species. The cDNA-sequence could be localized to human chromosome 11. In human cells the cDNA sequence is part of two large mRNAs. 4 different cosmid clones containing high molecular genomic DNA from normal human cells could be isolated from a library, each hybridizing to the AT500-cDNA. After stable transfection into AT-cells, one cosmid-clone was able to confer enhanced radiation resistance both in X-ray selections and dose-response experiments. The results indicate that the cloned cDNA-fragment is based on an unknown gene from human chromosome 11 which partially complements the radiosensitivity and the defective cell cycle progression in AT5BIVA cells. (orig.) [de

Evaluation of Antibacterial Activities of Citrus limon, Citrus reticulata, and Citrus grandis Against Pathogenic Bacteria

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Sholeh Saeb

2016-11-01

Full Text Available Background: Microorganisms resistant to most antibiotics are rapidly spreading, and there is an urgent and continuous need for novel antimicrobial compounds. The genus Citrus belongs to the family Rutaceae has many biologically active secondary metabolites. Objectives: The purpose of this study was to evaluate antimicrobial activity of essential oil and extract of Lemon (Citrus limon, Mandarin (Citrus reticulata and Pummelo (Citrus grandis against Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Salmonella typhi. Materials and Methods: The fresh Citrus leaves were shade-dried and powdered. Antimicrobial metabolites were extracted from them by 80% methanol for extract and using a Clevenger-type apparatus for essential oil. Eight different concentrations of the each leaf extract and essential oil were prepared. The antimicrobial susceptibility assay of Citrus leaves metabolites were subjected against four bacterial strains by agar disc diffusion and E-test method. Results: In this study, minimum inhibitory concentrations (MIC of different Citrus leaf extracts were determined against all four food-borne pathogens. The C. grandis leaf essential oil had potent antimicrobial activity against all four pathogens, and the C. limon leaf essential oil was effective on Gram-positive bacteria. S. typhi was resistant against two leaves essential oils. Conclusions: The results showed that there was no antimicrobial activity effect in all extracts on tested bacteria. In this study, the antibacterial effect of essential oil of Citrus leaves on four strains of pathogenic microorganisms was confirmed. The C. grandis leaf essential oil had the most powerful antimicrobial properties, suggesting its potential application as natural preservative in foods or an effective medicine against different pathogenic microbes. Key words: Antibacterial activity, E-test, Citr

Human thyroid peroxidase: complete cDNA and protein sequence, chromosome mapping, and identification of two alternately spliced mRNAs

International Nuclear Information System (INIS)

Kimura, S.; Kotani, T.; McBride, O.W.; Umeki, K.; Hirai, K.; Nakayama, T.; Ohtaki, S.

1987-01-01

Two forms of human thyroid peroxidase cDNAs were isolated from a λgt11 cDNA library, prepared from Graves disease thyroid tissue mRNA, by use of oligonucleotides. The longest complete cDNA, designated phTPO-1, has 3048 nucleotides and an open reading frame consisting of 933 amino acids, which would encode a protein with a molecular weight of 103,026. Five potential asparagine-linked glycosylation sites are found in the deduced amino acid sequence. The second peroxidase cDNA, designated phTPO-2, is almost identical to phTPO-1 beginning 605 base pairs downstream except that it contains 1-base-pair difference and lacks 171 base pairs in the middle of the sequence. This results in a loss of 57 amino acids corresponding to a molecular weight of 6282. Interestingly, this 171-nucleotide sequence has GT and AG at its 5' and 3' boundaries, respectively, that are in good agreement with donor and acceptor splice site consensus sequences. Using specific oligonucleotide probes for the mRNAs derived from the cDNA sequences hTOP-1 and hTOP-2, the authors show that both are expressed in all thyroid tissues examined and the relative level of two mRNAs is different in each sample. The results suggest that two thyroid peroxidase proteins might be generated through alternate splicing of the same gene. By using somatic cell hybrid lines, the thyroid peroxidase gene was mapped to the short arm of human chromosome 2

Enzyme characterisation, isolation and cDNA cloning of polyphenol oxidase in the hearts of palm of three commercially important species.

Science.gov (United States)

Shimizu, Milton Massao; Melo, Geraldo Aclécio; Brombini Dos Santos, Adriana; Bottcher, Alexandra; Cesarino, Igor; Araújo, Pedro; Magalhães Silva Moura, Jullyana Cristina; Mazzafera, Paulo

2011-09-01

Heart of palm (palmito) is the edible part of the apical meristem of palms and is considered a gourmet vegetable. Palmitos from the palms Euterpe edulis (Juçara) and Euterpe oleracea (Açaí) oxidise after harvesting, whereas almost no oxidation is observed in palmitos from Bactris gasipaes (Pupunha). Previous investigations showed that oxidation in Juçara and Açaí was mainly attributable to polyphenol oxidase (PPO; EC 1.14.18.1) activity. In this study, we partially purified PPOs from these three palmitos and analysed them for SDS activation, substrate specificity, inhibition by specific inhibitors, thermal stability, optimum pH and temperature conditions, Km and Ki. In addition, the total phenolic content and chlorogenic acid content were determined. Two partial cDNA sequences were isolated and sequenced from Açaí (EoPPO1) and Juçara (EePPO1). Semi-quantitative RT-PCR expression assays showed that Açaí and Juçara PPOs were strongly expressed in palmitos and weakly expressed in leaves. No amplification was observed for Pupunha samples. The lack of oxidation in the palmito Pupunha might be explained by the low PPO expression, low enzyme activity or the phenolic profile, particularly the low content of chlorogenic acid. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

Nucleotide sequence of a human cDNA encoding a ras-related protein (rap1B)

Energy Technology Data Exchange (ETDEWEB)

Pizon, V; Lerosey, I; Chardin, P; Tavitian, A [INSERM, Paris (France)

1988-08-11

The authors have previously characterized two human ras-related genes rap1 and rap2. Using the rap1 clone as probe they isolated and sequenced a new rap cDNA encoding the 184aa rap1B protein. The rap1B protein is 95% identical to rap1 and shares several properties with the ras protein suggesting that it could bind GTP/GDP and have a membrane location. As for rap1, the structural characteristics of rap1B suggest that the rap and ras proteins might interact on the same effector.

Nucleotide sequence of cloned cDNA for human sphingolipid activator protein 1 precursor

International Nuclear Information System (INIS)

Dewji, N.N.; Wenger, D.A.; O'Brien, J.S.

1987-01-01

Two cDNA clones encoding prepro-sphingolipid activator protein 1 (SAP-1) were isolated from a λ gt11 human hepatoma expression library using polyclonal antibodies. These had inserts of ≅ 2 kilobases (λ-S-1.2 and λ-S-1.3) and both were both homologous with a previously isolated clone (λ-S-1.1) for mature SAP-1. The authors report here the nucleotide sequence of the longer two EcoRI fragments of S-1.2 and S-1.3 that were not the same and the derived amino acid sequences of mature SAP-1 and its prepro form. The open reading frame encodes 19 amino acids, which are colinear with the amino-terminal sequence of mature SAP-1, and extends far beyond the predicted carboxyl terminus of mature SAP-1, indicating extensive carboxyl-terminal processing. The nucleotide sequence of cDNA encoding prepro-SAP-1 includes 1449 bases from the assigned initiation codon ATG at base-pair 472 to the stop codon TGA at base-pair 1921. The first 23 amino acids coded after the initiation ATG are characteristic of a signal peptide. The calculated molecular mass for a polypeptide encoded by 1449 bases is ≅ 53 kDa, in keeping with the reported value for pro-SAP-1. The data indicate that after removal of the signal peptide mature SAP-1 is generated by removing an additional 7 amino acids from the amino terminus and ≅ 373 amino acids from the carboxyl terminus. One potential glycosylation site was previously found in mature SAP-1. Three additional potential glycosylation sites are present in the processed carboxyl-terminal polypeptide, which they designate as P-2

Molecular cloning of chicken metallothionein. Deduction of the complete amino acid sequence and analysis of expression using cloned cDNA

Energy Technology Data Exchange (ETDEWEB)

Wei, D; Andrews, G K

1988-01-25

A cDNA library was constructed using RNA isolated from the livers of chickens which had been treated with zinc. This library was screened with a RNA probe complementary to mouse metallothionein-I (MT), and eight chicken MT cDNA clones were obtained. All of the cDNA clones contained nucleotide sequences homologous to regions of the longest (375 bp) cDNA clone. The latter contained an open reading frame of 189 bp, and the deduced amino acid sequence indicates a protein of 63 amino acids of which 20 are cysteine residues. Amino acid composition and partial amino acid sequence analyses of purified chicken MT protein agreed with the amino acid composition and sequence deduced from the cloned cDNA. Amino acid sequence comparison establish that chicken MT shares extensive homology with mammalian MTs. Southern blot analysis of chicken DNA indicates that the chicken MT gene is not a part of a large family of related sequences, but rather is likely to be a unique gene sequence. In the chicken liver, levels of chicken MT mRNA were rapidly induced by metals (Cd/sup 2 +/, Zn/sup 2 +/, Cu/sup 2 +/), glucocorticoids and lipopolysaccharide. MT mRNA was present in low levels in embryonic liver and increased to high levels during the first week after hatching before decreasing again to the basal levels found in adult liver. The results of this study establish that MT is highly conserved between birds and mammals and is regulated in the chicken by agents which also regulate expression of mammalian MT genes. However, in contrast to the mammals, the results suggest the existence of a single isoform of MT in the chicken.

Cloning of cDNA sequences of a progestin-regulated mRNA from MCF7 human breast cancer cells

Energy Technology Data Exchange (ETDEWEB)

Chalbos, D; Westley, B; Alibert, C; Rochefort, H

1986-01-24

A cDNA clone corresponding to an mRNA regulated by the progestin R5020, has been isolated by differential screening of a cDNA library from the MCF7 breast cancer cell line, which contains estrogen and progesterone receptors. This probe hybridized with a single species of poly A + RNA of 8-kb molecular weight as shown by Northern blot analysis and could also be used to total RNA preparation. This recombinant cone hybridized specifically to an mRNA coding for a 250,000 daltons protein when translated in vitro. This protein was identical to the 250 kDa progestin-regulated protein that the authors previously described as shown by immunoprecipitation with specific rabbit polyclonal antibodies. Dose-response curve and specificity studies show that the accumulation of the Pg8 mRNA and that of the 250-kDa protein was increased by 5 to 30-fold following progestin treatment and that this effect was mediated by the progesterone receptor. Time course of induction indicated that the accumulation of mRNA was rapid and preceded that of the protein. This is the first report on a cloned cDNA probe of progestin-regulated mRNA in human cell lines.

Molecular cloning of cDNA for the human tumor-associated antigen CO-029 and identification of related transmembrane antigens

International Nuclear Information System (INIS)

Szala, S.; Kasai, Yasushi; Steplewski, Z.; Rodeck, U.; Koprowski, H.; Linnenbach, A.J.

1990-01-01

The human tumor-associated antigen CO-029 is a monoclonal antibody-defined cell surface glycoprotein of 27-34 kDa. By using the high-efficiency COS cell expression system, a full-length cDNA clone for CO-029 was isolated. When transiently expressed in COS cells, the cDNA clone directed the synthesis of an antigen reactive to monoclonal antibody CO-029 in mixed hemadsorption and immunoblot assays. Sequence analysis revealed that CO-029 belongs to a family of cell surface antigens that includes the melanoma-associated antigen ME491, the leukocyte cell surface antigen CD37, and the Sm23 antigen of the parasitic helminth Schistosoma mansoni. CO-029 and ME491 antigen expression and the effect of their corresponding monoclonal antibodies on cell growth were compared in human tumor cell lines of various histologic origins

Development of three full-length infectious cDNA clones of distinct brassica yellows virus genotypes for agrobacterium-mediated inoculation.

Science.gov (United States)

Zhang, Xiao-Yan; Dong, Shu-Wei; Xiang, Hai-Ying; Chen, Xiang-Ru; Li, Da-Wei; Yu, Jia-Lin; Han, Cheng-Gui

2015-02-02

Brassica yellows virus is a newly identified species in the genus of Polerovirus within the family Luteoviridae. Brassica yellows virus (BrYV) is prevalently distributed throughout Mainland China and South Korea, is an important virus infecting cruciferous crops. Based on six BrYV genomic sequences of isolates from oilseed rape, rutabaga, radish, and cabbage, three genotypes, BrYV-A, BrYV-B, and BrYV-C, exist, which mainly differ in the 5' terminal half of the genome. BrYV is an aphid-transmitted and phloem-limited virus. The use of infectious cDNA clones is an alternative means of infecting plants that allows reverse genetic studies to be performed. In this study, full-length cDNA clones of BrYV-A, recombinant BrYV5B3A, and BrYV-C were constructed under control of the cauliflower mosaic virus 35S promoter. An agrobacterium-mediated inoculation system of Nicotiana benthamiana was developed using these cDNA clones. Three days after infiltration with full-length BrYV cDNA clones, necrotic symptoms were observed in the inoculated leaves of N. benthamiana; however, no obvious symptoms appeared in the upper leaves. Reverse transcription-PCR (RT-PCR) and western blot detection of samples from the upper leaves showed that the maximum infection efficiency of BrYVs could reach 100%. The infectivity of the BrYV-A, BrYV-5B3A, and BrYV-C cDNA clones was further confirmed by northern hybridization. The system developed here will be useful for further studies of BrYV, such as host range, pathogenicity, viral gene functions, and plant-virus-vector interactions, and especially for discerning the differences among the three genotypes. Copyright © 2014 Elsevier B.V. All rights reserved.

Large-scale transcriptional profiling of lignified tissues in Tectona grandis.

Science.gov (United States)

Galeano, Esteban; Vasconcelos, Tarcísio Sales; Vidal, Mabel; Mejia-Guerra, Maria Katherine; Carrer, Helaine

2015-09-15

Currently, Tectona grandis is one of the most valuable trees in the world and no transcript dataset related to secondary xylem is available. Considering how important the secondary xylem and sapwood transition from young to mature trees is, little is known about the expression differences between those successional processes and which transcription factors could regulate lignin biosynthesis in this tropical tree. Although MYB transcription factors are one of the largest superfamilies in plants related to secondary metabolism, it has not yet been characterized in teak. These results will open new perspectives for studies of diversity, ecology, breeding and genomic programs aiming to understand deeply the biology of this species. We present a widely expressed gene catalog for T. grandis using Illumina technology and the de novo assembly. A total of 462,260 transcripts were obtained, with 1,502 and 931 genes differentially expressed for stem and branch secondary xylem, respectively, during age transition. Analysis of stem and branch secondary xylem indicates substantial similarity in gene ontologies including carbohydrate enzymes, response to stress, protein binding, and allowed us to find transcription factors and heat-shock proteins differentially expressed. TgMYB1 displays a MYB domain and a predicted coiled-coil (CC) domain, while TgMYB2, TgMYB3 and TgMYB4 showed R2R3-MYB domain and grouped with MYBs from several gymnosperms and flowering plants. TgMYB1, TgMYB4 and TgCES presented higher expression in mature secondary xylem, in contrast with TgMYB2, TgHsp1, TgHsp2, TgHsp3, and TgBi whose expression is higher in young lignified tissues. TgMYB3 is expressed at lower level in secondary xylem. Expression patterns of MYB transcription factors and heat-shock proteins in lignified tissues are dissimilar when tree development was evaluated, obtaining more expression of TgMYB1 and TgMYB4 in lignified tissues of 60-year-old trees, and more expression in TgHsp1, TgHsp2, Tg

Migration and dispersal of Anthonomus grandis (Coleoptera: Curculionidae in South America

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Teodoro STADLER

2007-01-01

Full Text Available El presente estudio sobre la dispersión de Anthonomus grandis Boheman, el picudo del algodonero, en Argentina, Brasil, Paraguay y Bolivia, explora las características ecológicas y fisiológicas que han permitido a este insecto dispersarse y establecerse exitosamente en América del Sur. La plasticidad fenotípica de A. grandis se caracteriza por un tiempo de desarrollo flexible, ciclo de vida multivoltino con generaciones superpuestas, la capacidad de alimentarse con polen de diversas familias botánicas así como de otras fuentes de alimento y por su habilidad para migrar y dispersarse con la ayuda del viento. Todo esto hace de esta especie una plaga clave para el cultivo del algodón. Los cultivos de cítricos en Misiones, Argentina, son posibles sitios para la hibernación de esta especie. En esta región fueron capturadas grandes cantidades de individuos prediapausantes, provenientes de algodonales en post-cosecha en Paraguay, atraídos probablemente por compuestos volátiles de cítricos cultivados en la zona. La quiescencia facultativa que atraviesan los adultos ante condiciones adversas, conlleva a un retraso en el desarrollo que se relaciona con las condiciones desfavorables. Esto sugiere que la hibernación en A. grandis puede ser definida como «oligopausa», una forma intermedia de diapausa. Desde su introducción en Brasil en 1983 y hasta el 2006, el picudo se ha dispersado en dirección sudoeste hacia Argentina, a una velocidad promedio de 61 km año-1. Sin embargo, le ha insumido aproximadamente diez años cruzar 250 km, desde Paraguay hacia el centro de la zona algodonera de Argentina. Este progreso más lento se debe probablemente a las acciones llevadas a cabo en el marco del programa de erradicación del picudo del algodonero, por parte del gobierno de Argentina. La llegada del picudo al área central de cultivo de algodón en la Argentina, así como a otras áreas de cultivo en Paraguay y Argentina, confirma el hecho de que

Propriedades de chapas de flocos fabricadas com adesivo de uréia-formaldeído e de taninos da casca de Eucalyptus grandis W. Hill ex Maiden ou de Eucalyptus pellita F. Muell. Properties of flakeboards made from urea-formaldehyde and bark tannins adhesives of Eucalyptus grandis or Eucalyptus pellita

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Angélica de Cássia Oliveira Carneiro

2004-10-01

Full Text Available Os taninos foram extraídos da casca de Eucalyptus grandis e Eucalyptus pellita, com água quente, à qual se adicionaram 4,5% de sulfito de sódio, durante três horas. As temperaturas da solução foram iguais a 70 e 100 ºC para Eucalyptus grandis e Eucalyptus pellita, respectivamente. Para a produção dos adesivos e com o intuito de reduzir a sua viscosidade, os taninos foram sulfitados com sulfito de sódio e ácido acético. Formulações adesivas foram preparadas adicionando-se 0, 25, 50, 75 ou 100% de adesivos tânicos ao adesivo comercial de uréia-formaldeído. Foram fabricadas chapas de flocos de Pinus elliottii Engelm. e Eucalyptus grandis, utilizando-se 8% da formulação adesiva. As propriedades das chapas foram determinadas segundo a norma ASTM D-1037, de 1993. Observou-se que as propriedades das chapas foram superiores ao mínimo estabelecido pela norma ANSI/A 280.1-93, exceto no caso da resistência à umidade. Verificou-se, ainda, que o emprego de uma formulação adesiva contendo resina à base de uréia-formaldeído e tanino-formaldeído pode melhorar algumas propriedades.Bark tannins of Eucalyptus grandis and Eucalyptus pellita were extracted with 4,5% sodium sulfite in hot water solution for a period of three hours. Solution temperatures were 70 and 100ºC, for Eucalyptus grandis and Eucalyptus pellita bark respectively. Tannins were reacted with acetic acid and sodium sulfite to reduce adhesive viscosity. Adhesive formulations were prepared adding 0, 25, 50, 75 or 100% of tannin adhesives to the commercial urea-formaldehyde adhesive. Flakeboards were fabricated with 8% resin content. Board properties were determined according to ASTM D-1037 standards. Except for humidity, all board properties were superior to the values established by ANSI.A 208.1-93 commercial standard. Addition of tannins to the urea-formaldehyde adhesive improved some properties.

Cloning and expression of human deoxycytidine kinase cDNA

International Nuclear Information System (INIS)

Chottiner, E.G.; Shewach, D.S.; Datta, N.S.; Ashcraft, E.; Gribbin, D.; Ginsburg, D.; Fox, I.H.; Mitchell, B.S.

1991-01-01

Deoxycytidine (dCyd) kinase is required for the phosphorylation of several deoxyribonucleosides and certain nucleoside analogs widely employed as antiviral and chemotherapeutic agents. Detailed analysis of this enzyme has been limited, however, by its low abundance and instability. Using oligonucleotides based on primary amino acid sequence derived from purified dCyd kinase, the authors have screened T-lymphoblast cDNA libraries and identified a cDNA sequence that encodes a 30.5-kDa protein corresponding to the subunit molecular mass of the purified protein. Expression of the cDNA in Escherichia coli results in a 40-fold increase in dCyd kinase activity over control levels. Northern blot analysis reveals a single 2.8-kilobase mRNA expressed in T lymphoblasts at 5- to 10-fold higher levels than in B lymphoblasts, and decreased dCyd kinase mRNA levels are present in T-lymphoblast cell lines resistant to arabinofuranosylcytosine and dideoxycytidine. These findings document that this cDNA encodes the T-lymphoblast dCyd kinase responsible for the phosphorylation of dAdo and dGuo as well as dCyd and arabinofuranosylcytosine

Isolation of cowpea genes conferring drought tolerance ...

African Journals Online (AJOL)

The main objective of this study was to identify and isolate the genes conferring drought tolerance in cowpea. A cDNA library enriched for cowpea genes expressed specifically during responses to drought was constructed. A procedure called suppression subtractive hybridisation (SSH) was successfully employed to obtain ...

RESISTÊNCIA BIOLÓGICA DA MADEIRA TRATADA DE Eucalyptus grandis E Eucalyptus cloeziana A FUNGOS APODRECEDORES EM ENSAIOS DE LABORATÓRIO

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Magnos Alan Vivian

2015-01-01

Full Text Available The present study aimed to evaluate the biological resistance of treated wood from Eucalyptus grandis and Eucalyptus cloeziana under the action of biodeteriorative organisms in laboratory testing. Thus, we used trees of Eucalyptus grandis and Eucalyptus cloeziana, both 16 years old, which was converted into planks and subjected to preservative treatment in an autoclave with chromate copper arsenate (CCA. Then, it was made the specimens for the conduct of accelerated decay test, as recommended by ASTM. From the results, it was observed for the fungus Trametes versicolor that the preservative treatment was effective in reducing the biological degradation of the wood of the two species, with reduced mass loss in 35.17 and 82.31% for wood Eucalyptus grandis and Eucalyptus cloeziana, respectively, as for the fungus Gloeophyllum trabeum mass loss was reduced by 6.79 and 96.65%, compared to the control. Based on the conditions of realization of the present study, it was observed that preservative treatment with CCA is effective in the increasing the biological resistance of the wood under the action of fungi Trametes versicolor and Gloeophyllum trabeum.

Red de coexpresión de 320 genes de Tectona grandis Relacionados con procesos de estrés abiótico y xilogénesis

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Vladimir Camel

2017-01-01

Full Text Available Tectona grandis es un árbol maderable de importancia económica en bosques tropicales y subtropicales. Mediante este estudio, se identificaron familias de factores de transcripción (FTs y genes codificantes para enzima, diferencialmente expresados en el xilema del tallo, implicados en la regulación de la respuesta a estrés abiótico y xilogénesis en T. grandis . Así, fue analizada la distribución evolutiva de 19 genes codificantes para FTs de T. grandis mediante análisis filogenéticos. También, fue utilizada la minería de bases de datos y publicaciones para identificar 320 genes de Arabidopsis thaliana (ortólogos a T. grandis como soporte experimental y predictivo. Como resultados, se encontraron FTs de las familias bZIP, MYB, NAC, ER, b HLH , NuY y genes que codifican enzimas. Así mismo, se logró analizar el interactoma de T. grandis encontrando correlaciones de Pearson significativas para genes que regulan vías metabólicas de fenilpropanoides y estrés abiótico. Además, la red de coexpresión reveló nodos y aristas entre los genes TgRAP1, TgMyB1, TgHSF1, TgMyB3, TgNAC1, TgTsiid1, TgLieTFs1, TgNuy3, TgRAP2 y TgNuy4 . En particular, los análisis de ontología génica mostraron 31 genes de respuesta a estrés abiótico, principalmente TgHShT1, TgHSF1 y TgHSF2 como correguladores. Además, se encontró que el regulador maestro TgNAC1 , está involucrado en la corregulación de otros factores de transcripción.

Nucleotide sequence of Phaseolus vulgaris L. alcohol dehydrogenase encoding cDNA and three-dimensional structure prediction of the deduced protein.

Science.gov (United States)

Amelia, Kassim; Khor, Chin Yin; Shah, Farida Habib; Bhore, Subhash J

2015-01-01

Common beans (Phaseolus vulgaris L.) are widely consumed as a source of proteins and natural products. However, its yield needs to be increased. In line with the agenda of Phaseomics (an international consortium), work of expressed sequence tags (ESTs) generation from bean pods was initiated. Altogether, 5972 ESTs have been isolated. Alcohol dehydrogenase (AD) encoding gene cDNA was a noticeable transcript among the generated ESTs. This AD is an important enzyme; therefore, to understand more about it this study was undertaken. The objective of this study was to elucidate P. vulgaris L. AD (PvAD) gene cDNA sequence and to predict the three-dimensional (3D) structure of deduced protein. positive and negative strands of the PvAD cDNA clone were sequenced using M13 forward and M13 reverse primers to elucidate the nucleotide sequence. Deduced PvAD cDNA and protein sequence was analyzed for their basic features using online bioinformatics tools. Sequence comparison was carried out using bl2seq program, and tree-view program was used to construct a phylogenetic tree. The secondary structures and 3D structure of PvAD protein were predicted by using the PHYRE automatic fold recognition server. The sequencing results analysis showed that PvAD cDNA is 1294 bp in length. It's open reading frame encodes for a protein that contains 371 amino acids. Deduced protein sequence analysis showed the presence of putative substrate binding, catalytic Zn binding, and NAD binding sites. Results indicate that the predicted 3D structure of PvAD protein is analogous to the experimentally determined crystal structure of s-nitrosoglutathione reductase from an Arabidopsis species. The 1294 bp long PvAD cDNA encodes for 371 amino acid long protein that contains conserved domains required for biological functions of AD. The predicted deduced PvAD protein's 3D structure reflects the analogy with the crystal structure of Arabidopsis thaliana s-nitrosoglutathione reductase. Further study is required

Investigation of ESEM/EDX to measure liquor penetration and diffusion in Eucalyptus grandis wood chips during kraft pulping

CSIR Research Space (South Africa)

Grzeskowiak, V

2011-01-01

Full Text Available Environmental scanning electron microscopy coupled with energy dispersive x-ray (ESEM/EDX) was optimised to measure the penetration and diffusion of cooking liquor into Eucalyptus grandis wood chips during kraft pulping. The moisture content...

Use of glass transition temperature for stabilization of board's cracks of Eucalyptus grandis

Directory of Open Access Journals (Sweden)

Fred W. Calonego

2010-09-01

Full Text Available The Eucalyptus grandis logs temperatures were determined and correlated with the board's cracks during steaming. Thermocouples were inserted in the logs center, registering their temperatures during steaming at 90"C. The logs were sawed and the board's cracks measured. It was concluded that: (1 the logistic S-shaped curve explains the logs temperature variation; (2 the logs with diameter of 20 to As temperaturas em toras de Eucalyptus grandis, durante a vaporização, foram determinadas e correlacionadas com as rachaduras das tábuas. Nos centros das toras foram inseridos termopares e registradas suas temperaturas durante a vaporização à 90"C. As toras foram desdobradas e as rachaduras das tábuas mensuradas. Concluiu-se que: (1 o modelo estatístico sigmoidal logístico explica a variação da temperatura nas toras; (2 as toras com 20 a <25, 25 a <30 e 30 a <35 cm de diâmetro apresentaram, respectivamente, 84,2"C, 73,1"C e 45,8"C ao final da vaporização; e (3 as rachaduras foramsignificativamente menores nas toras que atingiram a temperatura de transição vítrea.

Effects of an inducible aiiA gene on disease resistance in Eucalyptus urophylla × Eucalyptus grandis.

Science.gov (United States)

Ouyang, L J; Li, L M

2016-08-01

N-acyl-homoserine lactones (AHLs) are metabolites of mostly gram-negative bacteria and are critical signaling molecules in bacterial quorum-sensing systems. At threshold concentrations, AHLs can activate the expression of pathogenic genes and induce diseases. Therefore, reducing AHL concentrations is a key point of disease control in plants. AHL-lactonase, which is expressed by aiiA, is widespread in Bacillus sp and can hydrolyze AHLs. In the present study, we cloned aiiA from Bacillus subtilis by PCR. A plant expression vector of aiiA was constructed and name Pcam-PPP3-aiiA, in which expression of aiiA was controlled by the pathogen-inducible plant promoter PPP3. The recombinant plasmid was transferred into Eucalyptus × urophylla × E. grandis by an Agrobacterium-mediated transformation. PCR and Southern blotting showed that aiiA was successfully integrated into the E. urophylla × E. grandis genome and its expression was induced by Ralstonia solanacearum 12 h after inoculation, as shown by reverse transcription-PCR. The transcription efficacy of aiiA increased 43.88-, 30.65-, and 18.95-fold after inoculation with R. solanacearum, Erwinia carotovora ssp. zeae (Sabet) and Cylindrocladium quinqueseptatum, respectively as shown by RT-real-time PCR. Transgenic E.urophylla × E.grandis expressing the AIIA protein exhibited significantly enhanced disease resistance compared to non-transgenic plants by delaying the onset of wilting and reducing the disease index.

D22S15 - a fetal brain cDNA with BanII and SacI RFLP

Energy Technology Data Exchange (ETDEWEB)

Rouleau, G A; Kurnit, D M; Neve, R L; Bazanowsky, A; Patterson, D; Gusella, J F

1988-02-25

A .58 kb single copy EcoRI fragment was isolated from a human fetal brain cDNA library and cloned into pBR322. This fragment recognizes a two allele polymorphism when used to probe human genomic DNA digested with SacI. There are no constant bands. Additional polymorphisms recognized by BanII and Bsp1286 are in disequilibrium with the BanII polymorphism. It has been localized to chromosome 22 by somatic cell hybrid analysis and linkage analysis. Co-dominant segregation has been observed in 15 informative families.

5'-end sequences of budding yeast full-length cDNA clones and quality scores - Budding yeast cDNA sequencing project | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available east_seq_qual.zip File URL: ftp://ftp.biosciencedbc.jp/archive/yeast_cdna/LATEST/...yeast_seq_qual.zip File size: 59.9MB Simple search URL http://togodb.biosciencedbc.jp/togodb/view/budding_yeast_cdna

Human beta 2 chain of laminin (formerly S chain): cDNA cloning, chromosomal localization, and expression in carcinomas

DEFF Research Database (Denmark)

Wewer, U M; Gerecke, D R; Durkin, M E

1994-01-01

or other known laminin genes. Immunostaining showed that the beta 2 chain is localized to the smooth muscle basement membranes of the arteries, while the homologous beta 1 chain is confined to the subendothelial basement membranes. The beta 2 chain was found in the basement membranes of ovarian carcinomas......Overlapping cDNA clones that encode the full-length human laminin beta 2 chain, formerly called the S chain, were isolated. The cDNA of 5680 nt contains a 5391-nt open reading frame encoding 1797 amino acids. At the amino terminus is a 32-amino-acid signal peptide that is followed by the mature...... beta 2 chain polypeptide of 1765 amino acids with a calculated molecular mass of 192,389 Da. The human beta 2 chain is predicted to have all of the seven structural domains typical of the beta chains of laminin, including the short cysteine-rich alpha region. The amino acid sequence of human beta 2...

Subtractive cloning of cDNA from Aspergillus oryzae differentially regulated between solid-state culture and liquid (submerged) culture.

Science.gov (United States)

Akao, Takeshi; Gomi, Katsuya; Goto, Kuniyasu; Okazaki, Naoto; Akita, Osamu

2002-07-01

In solid-state cultures (SC), Aspergillus oryzae shows characteristics such as high-level production and secretion of enzymes and hyphal differentiation with asexual development which are absent in liquid (submerged) culture (LC). It was predicted that many of the genes involved in the characteristics of A. oryzae in SC are differentially expressed between SC and LC. We generated two subtracted cDNA libraries with bi-directional cDNA subtractive hybridizations to isolate and identify such genes. Among them, we identified genes upregulated in or specific to SC, such as the AOS ( A. oryzae SC-specific gene) series, and those downregulated or not expressed in SC, such as the AOL ( A. oryzae LC-specific) series. Sequencing analyses revealed that the AOS series and the AOL series contain genes encoding extra- and intracellular enzymes and transport proteins. However, half were functionally unclassified by nucleotide sequences. Also, by expression profile, the AOS series comprised two groups. These gene products' molecular functions and physiological roles in SC await further investigation.

Activity of a Brazilian strain of Bacillus thuringiensis israelensis against the cotton Boll Weevil Anthonomus grandis Boheman (Coleoptera: Tenebrionidae).

Science.gov (United States)

Monnerat, R; Martins, E; Praça, L; Dumas, V; Berry, C

2012-02-01

A Brazilian Bacillus thuringiensis subspecies israelensis, toxic to Diptera, including mosquitoes, was found also to show toxicity to the coleopteran boll weevil Anthonomus grandis Boheman at an equivalent level to that of the standard coleopteran-active B. thuringiensis subspecies tenebrionis T08017. Recombinant B. thuringiensis strains expressing the individual Cyt1Aa, Cry4Aa, Cry4Ba and Cry11Aa toxins from this strain were assessed to evaluate their potential contribution to the activity against A. grandis, either alone or in combination. Whilst individual toxins produced mortality, none was sufficiently potent to allow calculation of LC50 values. Combinations of toxins were unable to attain the same potency as the parental B. thuringiensis subsp. israelensis, suggesting a major role for other factors produced by this strain.

Systematics and evolution of the Meriones shawii/grandis complex (Rodentia, Gerbillinae) during the Late Quaternary in northwestern Africa: Exploring the role of environmental and anthropogenic changes

Science.gov (United States)

Stoetzel, Emmanuelle; Cornette, Raphaël; Lalis, Aude; Nicolas, Violaine; Cucchi, Thomas; Denys, Christiane

2017-05-01

Rodents of the Meriones shawii/grandis complex have been attested to in North Africa since the Middle Pleistocene and are abundant in archaeological sites. Today, they are widely spread and represent a major pest to local human populations. This complex, therefore, represents an accurate model for investigating the roles of climate change and human impact in shaping Quaternary faunal diversity and distribution. Many gray areas still exist regarding the systematics, ecology and geographical distribution of this complex, for both present and past populations. The purpose of this study is to compare modern genotyped and fossil Meriones specimens in order to 1) clarify the current systematics and distribution of the Meriones populations of the shawii/grandis complex, 2) document the taxonomic diversity in fossil Meriones from northwestern Africa, and 3) track their phenotypic and biogeographic evolution through time. To answer these questions we used geometric morphometrics on skulls (landmarks) and first upper molars (landmarks and sliding landmarks). We evidenced the existence of two morpho-groups within the M. shawii/grandis complex, with a clear geographic pattern (M. grandis in Morocco vs. M. shawii in Algeria and Tunisia). Currently only one morpho-group, attributed to M. grandis, seems to exist in Morocco, with a small overlap with M. shawii in the most eastern part of the country. However, according to fossil data, M. shawii was also present in Atlantic Morocco during the Late Pleistocene. We have also highlighted the impact of Holocene climate change and habitat anthropization on this arid adapted group. During the Middle Holocene, a major climatic event (last interglacial optimum) seems to have induced a demographic collapse in Moroccan populations and the disappearance of the shawii clade from Morocco (except in the most eastern areas). Both species then re-expanded, benefitting from the increasing aridity and the new ecological niche driven by agriculture

cDNA and deduced primary structure of basic phospholipase A2 with neurotoxic activity from the venom secretion of the Crotalus durissus collilineatus rattlesnake

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F.H.R. Fagundes

2010-03-01

Full Text Available To illustrate the construction of precursor complementary DNAs, we isolated mRNAs from whole venom samples. After reverse transcription polymerase chain reaction (RT-PCR, we amplified the cDNA coding for a neurotoxic protein, phospholipase A2 D49 (PLA2 D49, from the venom of Crotalus durissus collilineatus (Cdc PLA2. The cDNA encoding Cdc PLA2 from whole venom was sequenced. The deduced amino acid sequence of this cDNA has high overall sequence identity with the group II PLA2 protein family. Cdc PLA2 has 14 cysteine residues capable of forming seven disulfide bonds that characterize this group of PLA2 enzymes. Cdc PLA2 was isolated using conventional Sephadex G75 column chromatography and reverse-phase high performance liquid chromatography (RP-HPLC. The molecular mass was estimated using matrix-assisted laser desorption ionization-time-of-flight (MALDI-TOF mass spectrometry. We tested the neuromuscular blocking activities on chick biventer cervicis neuromuscular tissue. Phylogenetic analysis of Cdc PLA2 showed the existence of two lines of N6-PLA2, denominated F24 and S24. Apparently, the sequences of the New World’s N6-F24-PLA2 are similar to those of the agkistrodotoxin from the Asian genus Gloydius. The sequences of N6-S24-PLA2 are similar to the sequence of trimucrotoxin from the genus Protobothrops, found in the Old World.

The identification of specific cDNA clones from tall and dwarf rice plants

International Nuclear Information System (INIS)

Youssefian, S.; Kamada, I.; Sano, H.

1990-01-01

Full text: The use of dwarfing genes in rice breeding has proceeded for several years without a clear understanding of the genetic, hormonal and physiological mechanisms involved. This issue was addressed by focussing on the isolation of specific clones from tall- and dwarf-derived cDNA libraries. The materials used include near-isogenic lines of the tall rice cultivar 'Shiokari', differing at the DGWG or 'Tanginbozu' dwarfing gene loci. Also used were tall and dwarf 'Ginbozu' rice, the latter having been induced by treatment with 5-azacytidine, a potent demethylating agent. Subtractive and differential hybridisation have, to date, identified several candidate tall- and dwarf-specific clones. Their further characterisation is currently underway. (author)

Sequence of a cDNA encoding turtle high mobility group 1 protein.

Science.gov (United States)

Zheng, Jifang; Hu, Bi; Wu, Duansheng

2005-07-01

In order to understand sequence information about turtle HMG1 gene, a cDNA encoding HMG1 protein of the Chinese soft-shell turtle (Pelodiscus sinensis) was amplified by RT-PCR from kidney total RNA, and was cloned, sequenced and analyzed. The results revealed that the open reading frame (ORF) of turtle HMG1 cDNA is 606 bp long. The ORF codifies 202 amino acid residues, from which two DNA-binding domains and one polyacidic region are derived. The DNA-binding domains share higher amino acid identity with homologues sequences of chicken (96.5%) and mammalian (74%) than homologues sequence of rainbow trout (67%). The polyacidic region shows 84.6% amino acid homology with the equivalent region of chicken HMG1 cDNA. Turtle HMG1 protein contains 3 Cys residues located at completely conserved positions. Conservation in sequence and structure suggests that the functions of turtle HMG1 cDNA may be highly conserved during evolution. To our knowledge, this is the first report of HMG1 cDNA sequence in any reptilian.

Above-ground dry matter accumulation by Eucalyptus grandis and its relation to standard meteorological data

International Nuclear Information System (INIS)

Byrne, G.F.

1989-01-01

The relationship between the rate of increase of biomass in some stands of Eucalyptus grandis, growing near Coffs Harbour, N.S.W., Australia, is explored in terms of estimated evapotranspiration and radiation interception, and related to a similar previous study of Pinus radiata. A possible role of method of planting, site slope and site aspect in biomass increase is also discussed

Intercropping Acacia mangium stimulates AMF colonization and soil phosphatase activity in Eucalyptus grandis

OpenAIRE

Bini, Daniel; Santos, Cristiane Alcantara dos; Silva, Mylenne Calcciolari Pinheiro da; Bonfim, Joice Andrade; Cardoso, Elke Jurandy Bran Nogueira

2018-01-01

ABSTRACT: Arbuscular mycorrhizal fungi (AMF) are very important to plant nutrition, mostly in terms of acquisition of P and micronutrients. While Acacia mangium is closely associated with AMF throughout the whole cycle, Eucalyptus grandis presents this symbiosis primarily at the seedling stage. The aim of this study was to evaluate the dynamics of AMF in these two tree species in both pure and mixed plantations during the first 20 months after planting. We evaluated the abundance, richness an...

Traits and trade-offs in whole-tree hydraulic architecture along the vertical axis of Eucalyptus grandis.

Science.gov (United States)

Pfautsch, Sebastian; Aspinwall, Michael J; Drake, John E; Chacon-Doria, Larissa; Langelaan, Rob J A; Tissue, David T; Tjoelker, Mark G; Lens, Frederic

2018-01-25

Sapwood traits like vessel diameter and intervessel pit characteristics play key roles in maintaining hydraulic integrity of trees. Surprisingly little is known about how sapwood traits covary with tree height and how such trait-based variation could affect the efficiency of water transport in tall trees. This study presents a detailed analysis of structural and functional traits along the vertical axes of tall Eucalyptus grandis trees. To assess a wide range of anatomical and physiological traits, light and electron microscopy was used, as well as field measurements of tree architecture, water use, stem water potential and leaf area distribution. Strong apical dominance of water transport resulted in increased volumetric water supply per unit leaf area with tree height. This was realized by continued narrowing (from 250 to 20 µm) and an exponential increase in frequency (from 600 to 13 000 cm-2) of vessels towards the apex. The widest vessels were detected at least 4 m above the stem base, where they were associated with the thickest intervessel pit membranes. In addition, this study established the lower limit of pit membrane thickness in tall E. grandis at ~375 nm. This minimum thickness was maintained over a large distance in the upper stem, where vessel diameters continued to narrow. The analyses of xylem ultrastructure revealed complex, synchronized trait covariation and trade-offs with increasing height in E. grandis. Anatomical traits related to xylem vessels and those related to architecture of pit membranes were found to increase efficiency and apical dominance of water transport. This study underlines the importance of studying tree hydraulic functioning at organismal scale. Results presented here will improve understanding height-dependent structure-function patterns in tall trees. © The Author(s) 2018. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Effects of impregnation of Eucalyptus grandis wood with fire-retardant salt on the production and quality of its charcoal; Efeito da impregnacao da madeira de Eucalyptus grandis com sais ignifugos na producao e na qualidade do carvao

Energy Technology Data Exchange (ETDEWEB)

Carvalho, Ana Marcia Macedo Ladeira; Vital, Benedito Rocha; Gomide, Jose Livio; Della Lucia, Ricardo Marius; Leite, Helio Garcia [Vicosa Univ., MG (Brazil). Dept. de Engenharia Florestal

1998-12-31

The present work was carried out to investigate the effects of impregnating Eucalyptus grandis wood fire retardant salts on the production and quality of its charcoal. The wood used came from a commercial stand of 72-month old Eucalyptus grandis planted at an initial 3.0 x 2.0 m spacing, established in Vicosa, Minas Gerais, Brazil. This wood was transformed in chips, treated with some fire-retardant salts and carbonized. results were evaluated through multivariate analysis. The difference among the 34 treatments, evaluated on the wood characteristics, based on group analysis, and using the distance D{sup 2} of Mahalanobis and the Tocher method, showed distinct group and subgroup formations. Based on technological data, treatment with ammonium sulphate at 15% concentration was classified as being the most promising one. The use of canonic variables analysis showed that the treatment with ammonium sulphate at a concentration of 5% was the best, and was given preference because of its lower sulphur content. (author) 12 refs., 1 fig., 7 tabs.

Field response of predatorRhizophagus grandis to prey frass and synthetic attractants.

Science.gov (United States)

Wainhouse, D; Beech-Garwood, P A; Howell, R S; Kelly, D; Orozco, M P

1992-10-01

A lure based on the proportional composition of monoterpenes inD. micans larval frass and deployed in Theysohn slot traps was highly attractive toR. grandis released in the field. The relative response to frass and lure was consistent over a range of doses, and behavior close to traps baited with either lure or frass appeared to be similar. The monoterpenes, formulated with antioxidant, appear to be stable over several weeks when released from proprietary reservoir and wick "air fresheners." The lure may be of value in monitoring predator populations.

Microarray-based method for the parallel analysis of genotypes and expression profiles of wood-forming tissues in Eucalyptus grandis

CSIR Research Space (South Africa)

Barros, E

2009-05-01

Full Text Available of Eucalyptus grandis planting stock that exhibit preferred wood qualities is thus a priority of the South African forestry industry. The researchers used microarray-based DNA-amplified fragment length polymorphism (AFLP) analysis in combination with expression...

3G vector-primer plasmid for constructing full-length-enriched cDNA libraries.

Science.gov (United States)

Zheng, Dong; Zhou, Yanna; Zhang, Zidong; Li, Zaiyu; Liu, Xuedong

2008-09-01

We designed a 3G vector-primer plasmid for the generation of full-length-enriched complementary DNA (cDNA) libraries. By employing the terminal transferase activity of reverse transcriptase and the modified strand replacement method, this plasmid (assembled with a polydT end and a deoxyguanosine [dG] end) combines priming full-length cDNA strand synthesis and directional cDNA cloning. As a result, the number of steps involved in cDNA library preparation is decreased while simplifying downstream gene manipulation, sequencing, and subcloning. The 3G vector-primer plasmid method yields fully represented plasmid primed libraries that are equivalent to those made by the SMART (switching mechanism at 5' end of RNA transcript) approach.

Thalassiolins A-C: new marine-derived inhibitors of HIV cDNA integrase.

Science.gov (United States)

Rowley, David C; Hansen, Mark S T; Rhodes, Denise; Sotriffer, Christoph A; Ni, Haihong; McCammon, J Andrew; Bushman, Frederic D; Fenical, William

2002-11-01

Human immunodeficiency virus (HIV) replication requires integration of viral cDNA into the host genome, a process mediated by the viral enzyme integrase. We describe a new series of HIV integrase inhibitors, thalassiolins A-C (1-3), isolated from the Caribbean sea grass Thalassia testudinum. The thalassiolins are distinguished from other flavones previously studied by the substitution of a sulfated beta-D-glucose at the 7-position, a substituent that imparts increased potency against integrase in biochemical assays. The most active of these molecules, thalassiolin A (1), displays in vitro inhibition of the integrase catalyzed strand transfer reaction (IC50=0.4 microM) and an antiviral IC50 of 30 microM. Molecular modeling studies indicate a favorable binding mode is probable at the catalytic core domain of HIV-1 integrase.

Microbial biomass and activity in litter during the initial development of pure and mixed plantations of Eucalyptus grandis and Acacia mangium Biomassa e atividade microbiana da serapilheira durante o desenvolvimento inicial de plantios puros e mistos de Eucalyptus grandis e Acacia mangium

Directory of Open Access Journals (Sweden)

Daniel Bini

2013-02-01

Full Text Available Studies on microbial activity and biomass in forestry plantations often overlook the role of litter, typically focusing instead on soil nutrient contents to explain plant and microorganism development. However, since the litter is a significant source of recycled nutrients that affect nutrient dynamics in the soil, litter composition may be more strongly correlated with forest growth and development than soil nutrient contents. This study aimed to test this hypothesis by examining correlations between soil C, N, and P; litter C, N, P, lignin content, and polyphenol content; and microbial biomass and activity in pure and mixed second-rotation plantations of Eucalyptus grandis and Acacia mangium before and after senescent leaf drop. The numbers of cultivable fungi and bacteria were also estimated. All properties were correlated with litter C, N, P, lignin and polyphenols, and with soil C and N. We found higher microbial activity (CO2 evolution in litter than in soil. In the E. grandis monoculture before senescent leaf drop, microbial biomass C was 46 % higher in litter than in soil. After leaf drop, this difference decreased to 16 %. In A. mangium plantations, however, microbial biomass C was lower in litter than in soil both before and after leaf drop. Microbial biomass N of litter was approximately 94 % greater than that of the soil in summer and winter in all plantations. The number of cultivable fungi and bacteria increased after leaf drop, especially so in the litter. Fungi were also more abundant in the E. grandis litter. In general, the A. mangium monoculture was associated with higher levels of litter lignin and N, especially after leaf drop. In contrast, the polyphenol and C levels in E. grandis monoculture litter were higher after leaf drop. These properties were negatively correlated with total soil C and N. Litter in the mixed stands had lower C:N and C:P ratios and higher N, P, and C levels in the microbial biomass. This suggests more

Systematic chemical profiling of Citrus grandis 'Tomentosa' by ultra-fast liquid chromatography/diode-array detector/quadrupole time-of-flight tandem mass spectrometry.

Science.gov (United States)

Li, Pan-lin; Liu, Meng-hua; Hu, Jie-hui; Su, Wei-wei

2014-03-01

Citrus grandis 'Tomentosa', as the original plant of the traditional Chinese medicine "Huajuhong", has been used as antitussive and expectorant in clinic for thousands of years. The fruit epicarp and whole fruit of this plant were both literarily recorded and commonly used. In the present study, an ultra-fast liquid chromatography coupled with diode-array detection and quadrupole/time-of-flight mass spectrometry (UFLC-DAD-Q-TOF-MS/MS) based chemical profiling method was developed for rapid holistic quality evaluation of C. grandis 'Tomentosa', which laid basis for chemical comparison of two medicinal parts. As a result, forty-eight constituents, mainly belonging to flavonoids and coumarins, were unambiguously identified by comparison with reference standards and/or tentatively characterized by elucidating UV spectra, quasi-molecular ions and fragment ions referring to information available in literature. Both of the epicarp and whole fruit samples were rich in flavonoids and coumarins, but major flavonoids contents in whole fruit were significantly higher than in epicarp (P<0.5). The proposed method could be useful in quality control and standardization of C. grandis 'Tomentosa' raw materials and its products. Results obtained in this study will provide a basis for quality assessment and further study in vivo. Copyright © 2013 Elsevier B.V. All rights reserved.

Behavior of Eucalyptus grandis and E. cloeziana seedlings grown in arsenic-contaminated soil Comportamento de mudas de E. grandis E. eucalyptus cloeziana cultivadas em solo contaminado por arsênio

Directory of Open Access Journals (Sweden)

Roseli Freire Melo

2010-06-01

Full Text Available Arsenic has been considered the most poisonous inorganic soil pollutant to living creatures. For this reason, the interest in phytoremediation species has been increasing in the last years. Particularly for the State of Minas Gerais, where areas of former mining activities are prone to the occurrence of acid drainage, the demand is great for suitable species to be used in the revegetation and "cleaning" of As-polluted areas. This study was carried out to evaluate the potential of seedlings of Eucalyptus grandis (Hill Maiden and E. cloeziana F. Muell, for phytoremediation of As-polluted soils. Soil samples were incubated for a period of 15 days with different As (Na2HAsO4 doses (0, 50, 100, 200, and 400 mg dm-3. After 30 days of exposure the basal leaves of E. cloeziana plants exhibited purple spots with interveinal chlorosis, followed by necrosis and death of the apical bud at the 400 mg dm-3 dose. Increasing As doses in the soil reduced root and shoot dry matter, plant height and diameter in both species, although the reduction was more pronounced in E. cloeziana plants. In both species, As concentrations were highest in the root system; the highest root concentration was found in E. cloeziana plants (305.7 mg kg-1 resulting from a dose of 400 mg dm-3. The highest As accumulation was observed in E. grandis plants, which was confirmed as a species with potential for As phytoextraction, tending to accumulate As in the root system and stem.O arsênio (As tem sido considerado o poluente inorgânico de solo mais tóxico para os seres vivos, razão pela qual o interesse por espécies indicadoras e fitorremediadoras tem aumentado nos últimos anos. Particularmente para o Estado de Minas Gerais, que apresenta áreas remanescentes de atividade mineradora sujeitas à ocorrência de drenagem ácida, existe grande demanda por espécies com potencial para serem utilizadas na revegetação e "limpeza" de substratos contaminados por esse metaloide. Este

Funções de afilamento não segmentadas e segmentadas para Tectona grandis na região centro-sul matogrossense Non-segmented and segmented taper models to Tectona grandis in center-southern region of Mato Grosso

Directory of Open Access Journals (Sweden)

Sidney Fernando Caldeira

2012-12-01

Full Text Available O objetivo deste estudo foi comparar a acurácia dos modelos polinomiais não segmentados do quinto grau e de Hradetzky de 1976 e os segmentados de Max e Burkhart de1976 e Clark et al. de 1991, na estimativa dos diâmetros ao longo do fuste de Tectona grandis L.f., com ajustes para o conjunto total dos dados e por classe de diâmetro, em um povoamento com 16 anos, na região centro- sul matogrossense. A base de dados foi composta por 114 árvores, cubadas pela metodologia de Hohenadl modificada e distribuídas em classes de diâmetro. O ajuste dos modelos foi avaliado em função do coeficiente de determinação corrigido, erro padrão da estimativa e pela distribuição dos resíduos em porcentagem. A acuracidade dos modelos ao longo do fuste foi avaliada pelo desvio, desvio padrão das diferenças, somatório de quadrado dos resíduos relativos e a porcentagem dos resíduos. A Equação de Hradetzky foi a que apresentou o melhor ajuste para estimar os diâmetros ao longo do fuste de Tectona grandis tanto para o conjunto total dos dados quanto para as classes de diâmetro com os menores valores nas estatísticas auxiliares, exceto na Classe 3, onde a equação selecionada foi a de Clark et al. The aim of this study was to compare the accuracy of the non-segmented polynomial models of Fifth degree and Hradetzky from 1976 and the segmented models of Max and Burkhart from 1976 and Clark et al. from 1991, in estimating diameters along the stem of Tectona grandis L.f., with adjustments for the full set of data and by diameter class in a 16 years old stand in the center–southern region of Mato Grosso State, Brazil. The database consisted of 114 trees, scaled by the modified Hohenadl method and distributed in diameter classes. The models adjustments were evaluated according to the adjusted coefficient of determination, standard eError of estimate and distribution of residuals. The accuracy of the models along the stem was evaluated by deviation

In vivo antitussive activity of Coccinia grandis against irritant aerosol and sulfur dioxide-induced cough model in rodents

Directory of Open Access Journals (Sweden)

Shakti Prasad Pattanayak and Priyashree Sunita

2009-12-01

Full Text Available Coccinia grandis (Cucurbitaceae has extensively used to get relief from asthma and cough by the indigenous people of India. The antitussive effect of aerosols of two different concentrations (2.5%, 5% w/v of methanol extract of C. grandis fruits were tested by counting the numbers of coughs produced due to aerosols of citric acid, 10 min after exposing the male guinea pigs to aerosols of test solutions for 7 min. In another set of experiment methanol extract was investigated for its therapeutic efficacy on a cough model induced by sulfur dioxide gas in mice. The results showed significant reduction of cough number obtained in the presence of both concentrations of methanol extract as that of the prototype antitussive agent codeine phosphate. Also, methanol extract exhibited significant antitussive effect at 100, 200 and 400 mg/kg, per orally by inhibiting the cough by 20.57, 33.73 and 56.71% within 90 min of performing the experiment respectively.

Integrating remote sensing and ancillary data for regional ecosystem assessment: eucalyptus grandis agro-system in kwazulu-Natal, South Africa

CSIR Research Space (South Africa)

Cho, Moses A

2009-07-01

Full Text Available The study demonstrates that the integration of remote sensing and in situ data could be important in providing more accurate estimates of E. grandis state in KwaZulu Natal, South Africa compared to remote sensing models. Such modelling effort would...

Organogenesis and transient genetic transformation of the hybrid Eucalyptus grandis × Eucalyptus urophylla Organogênese e transformação genética transiente do híbrido Eucalyptus grandis × Eucalyptus urophylla

Directory of Open Access Journals (Sweden)

Giovana Bomfim de Alcantara

2011-04-01

Full Text Available The hybrid Eucalyptus grandis x Eucalyptus urophylla presents high levels of productivity and potential for use in paper, cellulose and fiber industries. The bud organogenesis from leaf explants of two clones of E. grandis × E. urophylla was studied in order to verify the effect of several factors: subculture duration on multiplication medium, type of explants, entire and half leaves: basal and apical portions, and duration of the culture on a regeneration medium. Differences in organogenic capacity of the two clones tested were observed. The explant most recommended for organogenesis is the basal section of the leaf collected from shoot clusters subcultured every 17 days. Moreover, the leaf explants must be transferred to a fresh bud induction medium every five days. This study also aimed at evaluating factors affecting the genetic transformation of leaf explants with the uidA gene, via co-cultivation with Agrobacterium tumefaciens, such as the pre-culture of the explants on a specific medium, the duration of their co-culture with the bacteria and the addition of acetosyringone to the culture media. The best conditions for the expression of the uidA gene were two days of pre-culture of the leaf tissues, three days of co-culture with the bacteria and the addition of acetosyringone in pre- and co-culture media.O híbrido Eucalyptus grandis x Eucalyptus urophylla apresenta alta produtividade e potencial para indústrias de papel, celulose e fibras. A organogênese de explantes foliares de dois clones de E. grandis × E. urophylla foi estudada para verificar fatores como tempo de repicagem das plântulas matrizes em meio de multiplicação; tipo de explantes, folhas inteiras e de meias-folhas (porções basais e apicais e dos dias que os explantes foliares permaneceram em meio de regeneração. Foram observadas diferenças na capacidade organogênica dos dois clones testados. A parte basal das folhas, coletadas de brotações repicadas a cada 17

Diel and seasonal movements by adult Sacramento pikeminnow (Ptychocheilus grandis) in the Eel River, northwestern California

Science.gov (United States)

Bret C. Harvey; Rodney J. Nakamoto

1999-01-01

Abstract - In late summer and fall, radio-tagged adult Sacramento pike-minnow (Ptychocheilus grandis) at three sites in the Eel River of northwestern California moved more at night than during the day. Fish moved up to 535 m at night and returned to their original positions the following morning. Adult Sacramento pikeminnow at all sites occupied only pools during the...

Effect of soil water availability on gas exchange in young trees of Eucalyptus grandis W. Hill ex Maiden

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María Sara Mejía de Tafur

2017-10-01

Full Text Available Two outdoor trials were conducted in the department of Valle del Cauca, Colombia, in contrasting climatic conditions, to evaluate the effect of soil water availability on the gas exchange of four elite genotypes of Eucalyptus grandis as follows: 28-3, 18-3, 24A-5 and 19-1, respectively. One trial was conducted on the campus of the Universidad Nacional de Colombia located in Palmira, at 994 m.a.s.l. with an average temperature of 23.5°C (maximum of 31°C, minimum of 18°C, and 76% relative humidity (RH. The second trial was conducted at the nursery of Carton de Colombia S.A., located in Restrepo, at 1450 m.a.s.l. with an average temperature 18°C (maximum of 26°C, minimum of 12°C and 80% RH. A split-plot design with four replicates was used. Treatments were field capacity (FC, ½ FC, ¼ FC, and soil waterlogging. At both locations, pot surfaces were covered with plastic sheeting to prevent the entrance of rainfall. Significant differences between water regimes, genotypes and locations occurred in photosynthesis rate, internal CO2 of the substomatal cavity (Ci, stomatal conductance, and transpiration rate, indicating that E. grandis has physiological defense mechanisms to drought stress such as stomata closure, evidenced by a decreasing in photosynthesis rate, stomatal conductance, and transpiration rate. A differential response was also observed between genotypes depending on the environment, indicating a genotype x environment interaction in terms of plant physiological response. Eucalyptus grandis appears to save water under water stress conditions by a decreasing in photosynthesis rate. Excess water is more limiting than water deficit.

Fractionation of organic substances from the South African Eucalyptus grandis biomass by a combination of hot water and mild alkaline treatments

CSIR Research Space (South Africa)

Johakimu, Jonas K

2015-09-01

Full Text Available An alternative way of fractionating lignocellulose biomass into its individual components, hemicelluloses, lignin and cellulose, was investigated. South African Eucalyptus grandis wood chips were fractionated using a combination of hot water...

An Improved Quick Method for the Isolation of Total RNA from Cotton ...

African Journals Online (AJOL)

David PANG

2011-11-02

Nov 2, 2011 ... in liquid nitrogen in a mortar and pestle and stored until. RNA isolation. ... our laboratory for microarray analysis, cDNA pyro- sequencing studies and construction ..... Economic and rapid method for extracting cotton genomic ...

Isolation, cDNA cloning, and structure-based functional characterization of oryctin, a hemolymph protein from the coconut rhinoceros beetle, Oryctes rhinoceros, as a novel serine protease inhibitor.

Science.gov (United States)

Horita, Shoichiro; Ishibashi, Jun; Nagata, Koji; Miyakawa, Takuya; Yamakawa, Minoru; Tanokura, Masaru

2010-09-24

We isolated oryctin, a 66-residue peptide, from the hemolymph of the coconut rhinoceros beetle Oryctes rhinoceros and cloned its cDNA. Oryctin is dissimilar to any other known peptides in amino acid sequence, and its function has been unknown. To reveal that function, we determined the solution structure of recombinant (13)C,(15)N-labeled oryctin by heteronuclear NMR spectroscopy. Oryctin exhibits a fold similar to that of Kazal-type serine protease inhibitors but has a unique additional C-terminal α-helix. We performed protease inhibition assays of oryctin against several bacterial and eukaryotic proteases. Oryctin does inhibit the following serine proteases: α-chymotrypsin, endopeptidase K, subtilisin Carlsberg, and leukocyte elastase, with K(i) values of 3.9 × 10(-10) m, 6.2 × 10(-10) m, 1.4 × 10(-9) m, and 1.2 × 10(-8) m, respectively. Although the target molecule of oryctin in the beetle hemolymph remains obscure, our results showed that oryctin is a novel single domain Kazal-type inhibitor and could play a key role in protecting against bacterial infections.

PCR-identification of a Nicotiana plumbaginifolia cDNA homologous to the high-affinity nitrate transporters of the crnA family.

Science.gov (United States)

Quesada, A; Krapp, A; Trueman, L J; Daniel-Vedele, F; Fernández, E; Forde, B G; Caboche, M

1997-05-01

A family of high-affinity nitrate transporters has been identified in Aspergillus nidulans and Chlamydomonas reinhardtii, and recently homologues of this family have been cloned from a higher plant (barley). Based on six of the peptide sequences most strongly conserved between the barley and C. reinhardtii polypeptides, a set of degenerate primers was designed to permit amplification of the corresponding genes from other plant species. The utility of these primers was demonstrated by RT-PCR with cDNA made from poly(A)+ RNA from barley, C. reinhardtii and Nicotiana plumbaginifolia. A PCR fragment amplified from N. plumbaginifolia was used as probe to isolate a full-length cDNA clone which encodes a protein, NRT2;1Np, that is closely related to the previously isolated crnA homologue from barley. Genomic Southern blots indicated that there are only 1 or 2 members of the Nrt2 gene family in N. plumbaginifolia. Northern blotting showed that the Nrt2 transcripts are most strongly expressed in roots. The effects of external treatments with different N sources showed that the regulation of the Nrt2 gene(s) is very similar to that reported for nitrate reductase and nitrite reductase genes: their expression was strongly induced by nitrate but was repressed when reduced forms of N were supplied to the roots.

Dietary sugars and proline influence biological parameters of adult Trissolcus grandis, an egg parasitoid of Sunn pest, Eurygaster integriceps

NARCIS (Netherlands)

Hajirajabi, Nafiseh; Fazel, Morteza Movahedi; Harvey, Jeffrey A.; Arbab, Abbass; Asgari, Shahriar

Parasitoids are important natural enemies that are used in the biological control of insect herbivore pests. The egg parasitoid Trissolcus grandis Thompson (Hym. Scelionidae) is a major enemy of the Sunn pest, Eurygaster integriceps Puton (Hem. Scutelleridae), which in turn is one of the most

Vigas de madeira laminada colada com laminas pre-classificadas de eucalyptus grandis

OpenAIRE

Grohmann, Sandra Zampieri

1998-01-01

Dissertação (Mestrado) - Universidade Federal de Santa Catarina, Centro Tecnologico Comprovar a viabilidade de realização e desempenho de vigas de madeira laminada colada de Eucalyptus grandis classificadas de acordo com sua rigidez cujas lâminas estão divididas em classes de resistência onde o parâmetro de comparação é o módulo de elasticidade longitudinal. Para isso caracterizou-se física e mecanicamente a madeira e fez-se a classificação das laminas em grupos de acordo com o módulo de e...

Karyotypic analysis of the cotton boll weevil, Anthonomus grandis Boheman.

Science.gov (United States)

McNally, L R; Beck, M L; Biggers, C J

2000-01-01

The diploid chromosome number of the cotton boll weevil, Anthonomus grandis Boheman, is 44. Both C- and N-banding techniques of mitotic cells demonstrated constitutive heterochromatin in the p arm of the eight largest chromosomes, the p arm of the X chromosome, and the centromeric region of autosomal groups A-D. Neither the y nor the group E autosomes appeared to contain constitutive heterochromatin. Supernumerary chromosomes were not found in the boll weevil. Restriction endonuclease banding of primary spermatocytes revealed a rod-shaped Xy tetrad in which the X and y were terminally associated. The p arm of the large, submetacentric X was C-band positive. While two of the autosomal tetrads were typically ring-shaped in primary spermatocytes, the remaining 19 autosomal tetrads were rod-shaped.

Antioxidant capacity and larvicidal activity of essential oil and extracts from Lippia grandis

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Evelyn Ivana T. Damasceno

2011-02-01

Full Text Available The leaves and thin branches of Lippia grandis Schauer, Verbenaceae, are used for flavoring of food in the Brazilian Amazon, as substitute for oregano. In this study the constituents of the essential oil were identified and the antioxidant capacity and larvicidal activity of the oil and methanol extract and its sub-fractions were evaluated. A sensory evaluation was determined in view of absence of toxicity. The oil showed a yield of 2.1% and its main constituents were thymol (45.8%, p-cymene (14.3%, γ-terpinene (10.5%, carvacrol (9.9% and thymol methyl ether (4.8%, totalizing 85%. The DPPH radical scavenging activity showed values for the EC50 between 9.0 and 130.5 µg mL-1 and the TEAC/ABTS values varied from 131.1 to 336.0 mg TE/g, indicating significant antioxidant activity for the plant. The total phenolic content ranged from 223.0 to 761.4 mg GAE/g, contributing to the antioxidant activity observed. The crude extracts inhibited the bleaching of β-carotene and the oil showed the greatest inhibition (42.5%. The oil (LgO, 7.6±2.4 µg mL-1 showed strong larvicidal activity against the brine shrimp bioassay. The sensory evaluation was highly satisfactory in comparison to oregano. The results are very promising for the use of L. grandis in seasoning and antioxidant products.

Cloning and sequencing of the cDNA encoding a core protein of the paired helical filament of Alzheimer's disease: Identification as the microtubule-associated protein tau

International Nuclear Information System (INIS)

Goedert, M.; Wischik, C.M.; Crowther, R.A.; Walker, J.E.; Klug, A.

1988-01-01

Screening of cDNA libraries prepared from the frontal cortex of an Alzheimer's disease patient and from fetal human brain has led to isolation of the cDNA for a core protein of the paired helical filament of Alzheimer's disease. The partial amino acid sequence of this core protein was used to design synthetic oligonucleotide probes. The cDNA encodes a protein of 352 amino acids that contains a characteristic amino acid repeat in its carboxyl-terminal half. This protein is highly homologous to the sequence of the mouse microtubule-associated protein tau and thus constitutes the human equivalent of mouse tau. RNA blot analysis indicates the presence of two major transcripts, 6 and 2 kilobases long, with a wide distribution in normal human brain. Tau protein mRNAs were found in normal amounts in the frontal cortex from patients with Alzheimer's disease. The proof that at least part of tau protein forms a component of the paired helical filament core opens the way to understanding the mode of formation of paired helical filaments and thus, ultimately, the pathogenesis of Alzheimer's disease

Isolation and expression of the Pneumocystis carinii dihydrofolate reductase gene

DEFF Research Database (Denmark)

Edman, J C; Edman, U; Cao, Mi-Mi

1989-01-01

Pneumocystis carinii dihydrofolate reductase (DHFR; 5,6,7,8-tetrahydrofolate: NADP+ oxidoreductase, EC 1.5.1.3) cDNA sequences have been isolated by their ability to confer trimethoprim resistance to Escherichia coli. Consistent with the recent conclusion that P. carinii is a member of the Fungi...

Seleção de isolados de Beauveria bassiana patogênicos ao bicudo-do-algodoeiro Selection of isolates of Beauveria bassiana pathogenic to cotton boll weevil

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Carlos Alberto Domingues da Silva

2001-02-01

Full Text Available Objetivou-se selecionar isolados de Beauveria bassiana (Balsamo Vüillemin provenientes de diferentes hospedeiros e regiões geográficas, patogênicos ao Anthonomus grandis, o bicudo-do-algodoeiro. Foram analisados 12 isolados em condições de laboratório. Os isolados obtidos originalmente de A. grandis foram pouco virulentos a essa praga. A mortalidade do bicudo teve início no segundo dia após a inoculação das suspensões fúngicas, variando de 15% a 83%, com TL50 entre 5,30 a 11,06 dias. Os isolados apresentaram variabilidade quanto à germinação dos conídios em meio de cultura artificial, e esta não se correlacionou com a patogenicidade. O isolado CG138 de B. bassiana destacou-se como um dos mais virulentos ao bicudo-do-algodoeiro.The objective of this work was to screen pathogenic isolates of Beauveria bassiana (Balsamo Vüillemin from different hosts and geographic regions, pathogenic to the cotton boll weevil. Twelve isolates were evaluated. Isolates originally obtained from boll weevil showed low pathogenicity against this host. Insect mortality began on the second day after inoculation by fungal suspension, ranging from 15% to 83%, with a TL50 of 5.30 to 11.06 days. Variability among isolates concerning conidial germination in artificial culture medium was verified, and TG50 did not correlate with pathogenicity. One of the most virulent isolates of B. bassiana against cotton boll weevil is CG138.

Brotación in vitro de yemas de teca (Tectona grandis L. f. Sprouting buds in vitro teak (Tectona grandis L. f.

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Fabiana Rojas Parajeles

2012-11-01

Full Text Available La teca (Tectona grandis L.f. es una especie maderable exótica introducida a Costa Rica para la siembra de plantaciones comercial por la alta demanda de su madera, su rápido crecimiento y su alta calidad. Inicialmente se utilizó la semilla como único material de siembra y la pobre calidad de muchas de las plantaciones impulsó el inicio de programas de mejora genética. La propagación clonal tomó mucha importancia en estos programas y el cultivo in vitro se convirtió en una herramienta valiosa para la propagación masiva de los árboles élites. Por lo anterior, este trabajo se enfocó en evaluar el efecto de varias concentraciones de dos reguladores de crecimiento, bencilaminopurina (BA y ácido indolacético (AIB, solos y en combinación, en la brotación de yemas dormantes y formación de callos de teca. Tanto el análisis estadístico como la observación visual mostraron que el tratamiento que consistió de 0,005 mg/l de AIA fue el mejor para incrementar la brotación de las yemas y para disminuir la formación de callo.Teak (Tectona grandis L. f. is an exotic timber species introduced to Costa Rica for commercial plantations due to the high demand for its wood, rapid growth and high-quality. Initially the seed was use as the only planting material, but the poor quality of many of the plantations resulted in the initiation of genetic improvement programs. The introduction of clonal propagation in these programs and the establishment of in vitro culture techniques became import tools for mass propagation of the selected elite trees. On the foregoing, this work focused on assessing the effect of several concentrations of two growth regulators, benzylaminopurine (BA and indole acetic acid (AIB, alone and in combination, in the budding of dormantes buds and callus formation of teak. Both the statistical analysis and the visual observation showed that the treatment consisted of 0.005 mg/l of IAA was the best to increase the budding of the

Isolation of stress responsive Psb A gene from rice (Oryza sativa l.) using differential display.

Science.gov (United States)

Tyagi, Aruna; Chandra, Arti

2006-08-01

Differential display (DD) experiments were performed on drought-tolerant rice (Oryza sativa L.) genotype N22 to identify both upregulated and downregulated partial cDNAs with respect to moisture stress. DNA polymorphism was detected between drought-stressed and control leaf tissues on the DD gels. A partial cDNA showing differential expression, with respect to moisture stress was isolated from the gel. Northern blotting analysis was performed using this cDNA as a probe and it was observed that mRNA corresponding to this transcript was accumulated to high level in rice leaves under water deficit stress. At the DNA sequence level, the partial cDNA showed homology with psb A gene encoding for Dl protein.

Peptidomics combined with cDNA library unravel the diversity of centipede venom

DEFF Research Database (Denmark)

Rong, Mingqiang; Yang, Shilong; Wen, Bo

2015-01-01

of centipede venom. In the present study, we use peptidomics combined with cDNA library to uncover the diversity of centipede Scolopendra subspinipes mutilans L. Koch. 192 peptides were identified by LC-MS/MS and 79 precursors were deduced by cDNA library. Surprisingly, the signal peptides of centipede toxins...

Isolation and characterization of a cDNA encoding (S)-cis-N-methylstylopine 14-hydroxylase from opium poppy, a key enzyme in sanguinarine biosynthesis.

Science.gov (United States)

Beaudoin, Guillaume A W; Facchini, Peter J

2013-02-15

Sanguinarine is a benzo[c]phenenthridine alkaloid with potent antimicrobial properties found commonly in plants of the Papaveraceae, including the roots of opium poppy (Papaver somniferum). Sanguinarine is formed from the central 1-benzylisoquinoline intermediate (S)-reticuline via the protoberberine alkaloid (S)-scoulerine, which undergoes five enzymatic oxidations and an N-methylation. The first four oxidations from (S)-scoulerine are catalyzed by cytochromes P450, whereas the final conversion involves a flavoprotein oxidase. All but one gene in the biosynthetic pathway from (S)-reticuline to sanguinarine has been identified. In this communication, we report the isolation and characterization of (S)-cis-N-methylstylopine 14-hydroxylase (MSH) from opium poppy based on the transcriptional induction in elicitor-treated cell suspension cultures and root-specific expression of the corresponding gene. Along with protopine 6-hydroxylase, which catalyzes the subsequent and penultimate step in sanguinarine biosynthesis, MSH is a member of the CYP82N subfamily of cytochromes P450. The full-length MSH cDNA was expressed in Saccharomyces cerevisiae and the recombinant microsomal protein was tested for enzymatic activity using 25 benzylisoquinoline alkaloids representing a wide range of structural subgroups. The only enzymatic substrates were the N-methylated protoberberine alkaloids N-methylstylopine and N-methylcanadine, which were converted to protopine and allocryptopine, respectively. Copyright © 2013. Published by Elsevier Inc.

Spatial distribution of adult Anthonomus grandis Boheman (Coleoptera: Curculionidae and buds with feeding punctures on conventional and Bt cottonDistribuição espacial de adultos e botões com orifício de alimentação de Anthonomus grandis Boheman (Coleoptera: Curculionidae em algodoeiro convencional e Bt

Directory of Open Access Journals (Sweden)

Paulo Rogerio Beltramin da Fonseca

2013-06-01

Full Text Available O conhecimento dos arranjos de dispersão para adultos e botões com orifício de alimentação de Anthonomus grandis em cultivares de algodoeiro é necessário para aperfeiçoar o monitoramento e controle da praga. Esta pesquisa teve por objetivo realizar análises probabilísticas dos padrões de distribuição espacial dos adultos e botões com orifícios de alimentação de A. grandis em duas cultivares de algodão Bt e não Bt. O estudo foi conduzido a campo em duas áreas experimentais, cada uma composta por 100 parcelas de sete linhas de sete metros de comprimento. Em 16 amostragens avaliaram-se cinco plantas por parcela através da contagem dos adultos e dos botões com orifício de alimentaçãoentre janeiro e maio de 2010. Foram calculados os índices de dispersão (razão variância/média, índice de Morisita e Expoente k da Distribuição Binomial Negativa e as distribuições teóricas de freqüência (Poisson, Binomial Negativa e Binomial Positiva. Não houve diferença estatística entre as cultivares avaliadas. A distribuição espacial dos adultos de A. grandis, nas cultivares Bt e não Bt, ajustou-se nos arranjos probabilísticos de distribuição binomial negativa (agregado e distribuição binomial positiva (uniforme, conforme os dias após a emergência do algodoeiro. As análises de dispersão para os botões com orifícios de alimentação nas culturas Bt e convencional mostraram os modelos espaciais de Poisson (aleatório, distribuição binomial negativa (agregado e distribuição binomial positiva (uniforme, em seqüência, durante o ciclo da cultura.Dispersion patterns of Anthonomus grandis adults and damaged squares with their feeding punctures are important to enhance pest monitoring and control on cotton. In this research we performed probabilistic analyses of the distribution patterns of adults and squares with feeding punctures of A. grandis on two cotton genotypes, Bt and non-Bt near isogenic lines. We conducted

Reverse transcription using random pentadecamer primers increases yield and quality of resulting cDNA

DEFF Research Database (Denmark)

Stangegaard, Michael; Dufva, I.H.; Dufva, Hans Martin

2006-01-01

oligonucleotides (pentadecamers) consistently, yielded at least 2 fold as much cDNA as did random hexamers using either-poly(A) RNA or an amplified version of messenger RNA (aRNA) as a template. The cDNA generated using pentadecamers did not differ in size distribution or the amount of incorporated label compared...... with cDNA generated with random hexamers. The increased efficiency of priming using random pentadecamers resulted in reverse transcription of > 80% of the template aRNA, while random hexamers induced reverse transcription of only 40% of the template aRNA. This suggests a better coverage...... that random pentadecamers can replace random hexamers in reverse transcription reactions on both poly(A) RNA and amplified RNA, resulting in higher cDNA yields and quality....

Selection of Reference Genes for qRT-PCR Analysis of Gene Expression in Stipa grandis during Environmental Stresses.

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Dongli Wan

Full Text Available Stipa grandis P. Smirn. is a dominant plant species in the typical steppe of the Xilingole Plateau of Inner Mongolia. Selection of suitable reference genes for the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR is important for gene expression analysis and research into the molecular mechanisms underlying the stress responses of S. grandis. In the present study, 15 candidate reference genes (EF1 beta, ACT, GAPDH, SamDC, CUL4, CAP, SNF2, SKIP1, SKIP5, SKIP11, UBC2, UBC15, UBC17, UCH, and HERC2 were evaluated for their stability as potential reference genes for qRT-PCR under different stresses. Four algorithms were used: GeNorm, NormFinder, BestKeeper, and RefFinder. The results showed that the most stable reference genes were different under different stress conditions: EF1beta and UBC15 during drought and salt stresses; ACT and GAPDH under heat stress; SKIP5 and UBC17 under cold stress; UBC15 and HERC2 under high pH stress; UBC2 and UBC15 under wounding stress; EF1beta and UBC17 under jasmonic acid treatment; UBC15 and CUL4 under abscisic acid treatment; and HERC2 and UBC17 under salicylic acid treatment. EF1beta and HERC2 were the most suitable genes for the global analysis of all samples. Furthermore, six target genes, SgPOD, SgPAL, SgLEA, SgLOX, SgHSP90 and SgPR1, were selected to validate the most and least stable reference genes under different treatments. Our results provide guidelines for reference gene selection for more accurate qRT-PCR quantification and will promote studies of gene expression in S. grandis subjected to environmental stress.

Blood Sugar Lowering Effect of Coccinia grandis (L. J. Voigt: Path for a New Drug for Diabetes Mellitus

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M. A. A. K. Munasinghe

2011-01-01

Full Text Available Background. Role of herbs in the management and control of diabetes has emerged fast over the years. We assessed the efficacy of Coccinia grandis (locally known as Ken, Kovakka leaves as a hypoglycemic agent. Methods. Double-blind phase I clinical trial was conducted at the general hospital and a private hospital in Matara in August 2009. All the participants were given a common meal for dinner, and they maintained a 10-hour fasting period. Sixty-one healthy volunteers were given a meal containing 20 g of leaves of Coccinia grandis which was mixed with a measured amount of scraped coconut and table salt for breakfast, and other 61 were given the placebo meal which also contained scraped coconut and salt. Glucose tolerance test was performed blindly for the two groups. Mixed factorial design analysis of variance and student's t-test were applied. Results. Overall blood sugar levels of the experimental group were also significantly lower than those of the control group (F(1,117 5.56, <0.05. Increase in the blood sugar levels from fasting to one hour (F(1,117 6.77, <0.05 and two hours (F(1,117 5.28, <0.05 postprandially was statistically significant for participants who were in the control group than those of in the experimental group. The mean difference of postprandial blood sugar levels (mg/dL after one hour (20.2, 95% confidence interval, 4.81 to 35.5 and two hours (11.46, 95% confidence interval; 1.03 to 21.9 was statistically significant between the two groups. Conclusions. Coccinia grandis has a blood sugar lowering effect. However further studies are needed to validate our findings.

Functional study and regional mapping of 44 hormono-regulated genes isolated from a porcine granulosa cell library

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Hatey François

2001-01-01

Full Text Available Abstract cDNA clones from a pig granulosa cell cDNA library were isolated by differential hybridisation for follicle stimulating hormone (FSH regulation in granulosa cells in a previous study. The clones that did not match any known sequence were studied for their expression in granulosa cells (treated or not by FSH and in fresh isolated ovarian follicles mainly by comparative RT-PCR analysis. These results give functional data on genes that may be implicated in follicular growing. These ESTs have been localised on the porcine genome, using a somatic cell hybrid panel, providing new type I markers on the porcine map and information on the comparative map between humans and pigs.

cDNA cloning of porcine brain prolyl endopeptidase and identification of the active-site seryl residue

Energy Technology Data Exchange (ETDEWEB)

Rennex, D.; Hemmings, B.A.; Hofsteenge, J.; Stone, S.R. (Friedrich Miescher-Institut, Basel (Switzerland))

1991-02-26

Prolyl endopeptidase is a cytoplasmic serine protease. The enzyme was purified from porcine kidney, and oligonucleotides based on peptide sequences from this protein were used to isolate a cDNA clone from a porcine brain library. This clone contained the complete coding sequence of prolyl endopeptidase and encoded a polypeptide with a molecular mass of 80751 Da. The deduced amino acid sequence of prolyl endopeptidase showed no sequence homology with other known serine proteases. ({sup 3}H)Diisopropyl fluorophosphate was used to identify the active-site serine of prolyl endopeptidase. One labeled peptide was isolated and sequenced. The sequence surrounding the active-site serine was Asn-Gly-Gly-Ser-Asn-Gly-Gly. This sequence is different from the active-site sequences of other known serine proteases. This difference and the lack of overall homology with the known families of serine proteases suggest that prolyl endopeptidase represents a new type of serine protease.

Examination of gene expression in mice exposed to low dose radiation using affymetrix cDNA microarrays

Energy Technology Data Exchange (ETDEWEB)

Morris, D.; Knox, D.; Lavoie, J.; Lemon, J.; Boreham, D. [McMaster Univ., Hamilton, Ontario (Canada)

2005-07-01

'Full text:' Gamma radiation acts via the indirect effect to damage cells by producing reactive oxygen species (ROS). These ROS are capable damaging macromolecules and, altering signal pathways and gene transcription. Cells have evolved enzymes and mechanisms to scavenge ROS and repair oxidative damage. Microarrays allow the survey of the gene transcription activity of thousands of genes simultaneously. Messenger RNA is extracted from cells, hybridized with the complementary DNA (cDNA) of a microarray chip, and examined with a chip reader. Affymetrix microarray chips have been produced by the CSCHAH in Winnipeg containing 26000 murine genes. Groups of female mice have been exposed to low dose whole body chronic gamma radiation exposures of 0,50,100, and 120 mGy, corresponding to 15,30,60, and 75 weeks, respectively. MRNA from mice brain tissue has been extracted, isolated, converted to cDNA and labeled. Gene expression in each irradiated mouse was compared to the pooled expression of the control mice. Analysis of gene expression levels are performed with microarray analytical software, Array Pro by Media Cybernetics, and powerful statistical software, BRB microarray tools. Differences in gene expressions, focusing on genes for cytokines, DNA repair mechanisms, immuno-modulators, apoptosis pathways, and enzymatic anti-oxidant systems, are being examined and will be reported. (author)

Qualidade de juntas coladas com lâminas de madeira oriundas de três regiões do tronco de Eucalyptus grandis, Eucalyptus saligna e Pinus elliottii Quality of wood joints glued with wood veneers from three trunk regions of Eucalyptus grandis, Eucalyptus saligna and Pinus elliottii

Directory of Open Access Journals (Sweden)

Benedito Rocha Vital

2006-08-01

Full Text Available Este experimento teve como objetivo avaliar a resistência de juntas coladas formadas pelas combinações de lâminas provenientes de três posições no tronco da madeira de Eucalyptus grandis, Eucalyptus saligna e Pinus elliottii. Foram empregados adesivos à base de poliacetato de vinila de média e alta viscosidade e resorcinol-formaldeído nas gramaturas de 150 g/m², em face simples para o poliacetato de vinila de média e alta viscosidades e 300 g/m², em face dupla, para o adesivo resorcinólico. O teor médio de umidade das lâminas, no momento da colagem, foi igual a 14%. Os valores médios mais elevados de resistência ao cisalhamento foram obtidos nas juntas produzidas com madeira de Eucalyptus saligna, coladas com adesivos de poliacetato de média viscosidade e resorcinol-formaldeído. A maior porcentagem de falha profunda na madeira foi obtida em juntas de madeira de Pinus elliottii, unidas com adesivo de poliacetato de alta viscosidade, seguidas das juntas de Eucalyptus grandis e coladas com adesivo de poliacetato de média viscosidade. As combinações de lâminas oriundas das seguintes posições no tronco: medula e casca, intermediária e casca e casca e casca resultaram em linhas de cola com maiores resistências ao cisalhamento.The objective of this work was to evaluate the shear strength of glued wood joints from pith, outer and intermediary wood of Eucalyptus saligna, Eucalyptus grandis and Pinus elliottii. High and medium viscosity polyvinyl acetate and resorcinol-phenol adhesives were applied at spread rate of 150 g/m² in single line and at spread rate of 300 g/m² in double glue line for the resorcinolic adhesive. The mean wood moisture content was 14%. Higher shear strength was obtained with Eucalyptus saligna veneer glued with medium viscosity polyvinyl resorcinolic adhesive. The highest percentage of wood failure was found on Pinus elliottii veneer glued with high viscosity polyvinyl acetate adhesive followed by

PEX12, the pathogenic gene of group III Zellweger syndrome: cDNA cloning by functional complementation on a CHO cell mutant, patient analysis, and characterization of PEX12p

NARCIS (Netherlands)

Okumoto, K.; Shimozawa, N.; Kawai, A.; Tamura, S.; Tsukamoto, T.; Osumi, T.; Moser, H.; Wanders, R. J.; Suzuki, Y.; Kondo, N.; Fujiki, Y.

1998-01-01

Rat PEX12 cDNA was isolated by functional complementation of peroxisome deficiency of a mutant CHO cell line, ZP109 (K. Okumoto, A. Bogaki, K. Tateishi, T. Tsukamoto, T. Osumi, N. Shimozawa, Y. Suzuki, T. Orii, and Y. Fujiki, Exp. Cell Res. 233:11-20, 1997), using a transient transfection assay and

MICOBIOTA ASSOCIADA À MADEIRA SERRADA DE Eucalyptus grandis HILL EX MAIDEN DURANTE A SECAGEM AO AR LIVRE

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Paulo Fernando Trugilho

2006-02-01

Full Text Available O objetivo principal deste trabalho foi o de acompanhar a secagem ao ar livre da madeira serrada de Eucalyptus grandis conduzida na região de Lavras, MG e identificar a micobiota associada à madeira em diversas fases da secagem. As tábuas foram cortadas de oito toras de 3,0 m de comprimento, oriundas de três árvores, com 27 anos de idade, plantadas em área experimental da UFLA. Dos resultados, pôde-se concluir que a secagem da madeira de Eucalyptus grandis ao ar livre, iniciada em janeiro, consumiu 158 dias até atingir umidade próxima a 12,5%. A curva de secagem foi representada por uma equação logarítmica com coeficiente de determinação (R2 igual a 98,3%. Os fungos causadores de bolores superficiais e de manchas tiveram maior ocorrência no início da secagem. As mais altas freqüências de fungos, verificadas no início da secagem, foram observadas para Penicillium spp. e Pestalotiopsis sp. O fungo Lentinus lepideus ocorreu com maior freqüência no final da secagem.

High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis

CSIR Research Space (South Africa)

Van den Berg, N

2004-11-01

Full Text Available Efficient construction of cDNA libraries enriched for differentially expressed transcripts is an important first step in many biological investigations. We present a quantitative procedure for screening cDNA libraries constructed by suppression...

Construction and identification of subtracted cDNA library in bone marrow cells of radon-exposed mice

International Nuclear Information System (INIS)

Li Jianxiang; Nie Jihua; Tong Jian; Fu Chunling; Zhou Jianwei

2008-01-01

Objective: To construct and identify subtracted cDNA library in bone marrow cells of mice exposed to radon inhalation. Methods: Adult male BALB/c mice, weighing 18-22 g, were placed in a multi- functional radon chamber. One group of mice was exposed to radon up to the accumulative dose of 105 work level month (WLM). The control group of mice was housed in a room with an accumulative dose of 1 WLM. To construct a subtracted cDNA library enriched with differentially expressed genes, the SMART technique and the suppression subtractive hybridization were performed. The obtained forward and reverse cDNA fragments were directly inserted into pMD18-T vector and transformed into E. coli JM109. The inserting cDNA fragments were screened by the blue-and-white blot screening and nested PCR of bacterium liquid. Results: The 244 of 285 white bacteria clones obtained randomly were positive clones contained 100-1100 bp inserted cDNA fragments. Conclusions: The forward and reverse subtracted cDNA library in bone marrow cells of mice exposed to radon inhalation is successfully constructed. (authors)

Microbial biomass and activity in litter during the initial development of pure and mixed plantations of Eucalyptus grandis and Acacia mangium

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Daniel Bini

2013-02-01

Full Text Available Studies on microbial activity and biomass in forestry plantations often overlook the role of litter, typically focusing instead on soil nutrient contents to explain plant and microorganism development. However, since the litter is a significant source of recycled nutrients that affect nutrient dynamics in the soil, litter composition may be more strongly correlated with forest growth and development than soil nutrient contents. This study aimed to test this hypothesis by examining correlations between soil C, N, and P; litter C, N, P, lignin content, and polyphenol content; and microbial biomass and activity in pure and mixed second-rotation plantations of Eucalyptus grandis and Acacia mangium before and after senescent leaf drop. The numbers of cultivable fungi and bacteria were also estimated. All properties were correlated with litter C, N, P, lignin and polyphenols, and with soil C and N. We found higher microbial activity (CO2 evolution in litter than in soil. In the E. grandis monoculture before senescent leaf drop, microbial biomass C was 46 % higher in litter than in soil. After leaf drop, this difference decreased to 16 %. In A. mangium plantations, however, microbial biomass C was lower in litter than in soil both before and after leaf drop. Microbial biomass N of litter was approximately 94 % greater than that of the soil in summer and winter in all plantations. The number of cultivable fungi and bacteria increased after leaf drop, especially so in the litter. Fungi were also more abundant in the E. grandis litter. In general, the A. mangium monoculture was associated with higher levels of litter lignin and N, especially after leaf drop. In contrast, the polyphenol and C levels in E. grandis monoculture litter were higher after leaf drop. These properties were negatively correlated with total soil C and N. Litter in the mixed stands had lower C:N and C:P ratios and higher N, P, and C levels in the microbial biomass. This suggests more

Migration and dispersal of Anthonomus grandis (Coleoptera: Curculionidae in South America Migración y dispersión de Anthonomus grandis (Coleoptera: Curculionidae en América del Sur

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Teodoro Stadler

2007-12-01

Full Text Available This study discusses the dispersal of Anthonomus grandis Boheman, the cotton boll weevil, in Argentina, Brazil, Paraguay and Bolivia, exploring the ecological and physiological factors that have made the dispersal and establishment of this insect in South America so successful. The boll weevil's phenotypic plasticity is represented by its flexible developmental time, its multivoltine life cycle with several overlapping generations, its capacity to feed on pollen from diverse botanical families as well as from non pollen food sources and its ability to migrate and disperse aided by winds. These characteristics make it a key pest for cotton. Probable overwintering «hot spots» for the boll weevil were identified in Misiones-Argentina, where large numbers of prediapausing weevils concentrate after arrival from newly harvested cotton fields in Paraguay, probably attracted by citrus orchards volatiles. The boll weevil's facultative quiescence is always relative to environmental adverse conditions. This suggests that overwintering in the boll weevil can be defined as «oligopause», an intermediate form of diapause. Since its introduction to Brazil in 1983, until 2006, it has spread southwest at an average of 61 km year-1 towards Argentina. However, it took the boll weevil approximately ten years to move 250 km between Paraguay and the main cotton growing area in Argentina. This slower progress is probably due to the actions taken by the Argentine government through the boll weevil eradication program. The arrival of the boll weevil at the cotton cropping areas in Paraguay and Argentina reinforces the fact that the boll weevil should finally be included in an integrated cotton pest management program jointly with other major cotton pests.El presente estudio sobre la dispersión de Anthonomus grandis Boheman, el picudo del algodonero, en Argentina, Brasil, Paraguay y Bolivia, explora las características ecológicas y fisiológicas que han permitido a

cDNA sequencing improves the detection of P53 missense mutations in colorectal cancer

International Nuclear Information System (INIS)

Szybka, Malgorzata; Kordek, Radzislaw; Zakrzewska, Magdalena; Rieske, Piotr; Pasz-Walczak, Grazyna; Kulczycka-Wojdala, Dominika; Zawlik, Izabela; Stawski, Robert; Jesionek-Kupnicka, Dorota; Liberski, Pawel P

2009-01-01

Recently published data showed discrepancies beteween P53 cDNA and DNA sequencing in glioblastomas. We hypothesised that similar discrepancies may be observed in other human cancers. To this end, we analyzed 23 colorectal cancers for P53 mutations and gene expression using both DNA and cDNA sequencing, real-time PCR and immunohistochemistry. We found P53 gene mutations in 16 cases (15 missense and 1 nonsense). Two of the 15 cases with missense mutations showed alterations based only on cDNA, and not DNA sequencing. Moreover, in 6 of the 15 cases with a cDNA mutation those mutations were difficult to detect in the DNA sequencing, so the results of DNA analysis alone could be misinterpreted if the cDNA sequencing results had not also been available. In all those 15 cases, we observed a higher ratio of the mutated to the wild type template by cDNA analysis, but not by the DNA analysis. Interestingly, a similar overexpression of P53 mRNA was present in samples with and without P53 mutations. In terms of colorectal cancer, those discrepancies might be explained under three conditions: 1, overexpression of mutated P53 mRNA in cancer cells as compared with normal cells; 2, a higher content of cells without P53 mutation (normal cells and cells showing K-RAS and/or APC but not P53 mutation) in samples presenting P53 mutation; 3, heterozygous or hemizygous mutations of P53 gene. Additionally, for heterozygous mutations unknown mechanism(s) causing selective overproduction of mutated allele should also be considered. Our data offer new clues for studying discrepancy in P53 cDNA and DNA sequencing analysis

Efficacy Testing of H56 cDNA Tattoo Immunization against Tuberculosis in a Mouse Model.

Science.gov (United States)

Platteel, Anouk C M; Nieuwenhuizen, Natalie E; Domaszewska, Teresa; Schürer, Stefanie; Zedler, Ulrike; Brinkmann, Volker; Sijts, Alice J A M; Kaufmann, Stefan H E

2017-01-01

Tuberculosis (TB), caused by Mycobacterium tuberculosis ( Mtb ), remains a global threat. The only approved vaccine against TB, Mycobacterium bovis bacillus Calmette-Guérin (BCG), provides insufficient protection and, being a live vaccine, can cause disseminated disease in immunocompromised individuals. Previously, we found that intradermal cDNA tattoo immunization with cDNA of tetanus toxoid fragment C domain 1 fused to cDNA of the fusion protein H56, comprising the Mtb antigens Ag85B, ESAT-6, and Rv2660c, induced antigen-specific CD8 + T cell responses in vivo . As cDNA tattoo immunization would be safer than a live vaccine in immunocompromised patients, we tested the protective efficacy of intradermal tattoo immunization against TB with H56 cDNA, as well as with H56_E, a construct optimized for epitope processing in a mouse model. As Mtb antigens can be used in combination with BCG to boost immune responses, we also tested the protective efficacy of heterologous prime-boost, using dermal tattoo immunization with H56_E cDNA to boost BCG immunization in mice. Dermal H56 and H56_E cDNA immunization induced H56-specific CD4 + and CD8 + T cell responses and Ag85B-specific IgG antibodies, but did not reduce bacterial loads, although immunization with H56_E ameliorated lung pathology. Both subcutaneous and intradermal immunization with BCG resulted in broad cellular immune responses, with increased frequencies of CD4 + T effector memory cells, T follicular helper cells, and germinal center B cells, and resulted in reduced bacterial loads and lung pathology. Heterologous vaccination with BCG/H56_E cDNA induced increased H56-specific CD4 + and CD8 + T cell cytokine responses compared to vaccination with BCG alone, and lung pathology was significantly decreased in BCG/H56_E cDNA immunized mice compared to unvaccinated controls. However, bacterial loads were not decreased after heterologous vaccination compared to BCG alone. CD4 + T cells responding to Ag85B- and ESAT-6

Isolation, cDNA Cloning, and Structure-based Functional Characterization of Oryctin, a Hemolymph Protein from the Coconut Rhinoceros Beetle, Oryctes rhinoceros, as a Novel Serine Protease Inhibitor*

Science.gov (United States)

Horita, Shoichiro; Ishibashi, Jun; Nagata, Koji; Miyakawa, Takuya; Yamakawa, Minoru; Tanokura, Masaru

2010-01-01

We isolated oryctin, a 66-residue peptide, from the hemolymph of the coconut rhinoceros beetle Oryctes rhinoceros and cloned its cDNA. Oryctin is dissimilar to any other known peptides in amino acid sequence, and its function has been unknown. To reveal that function, we determined the solution structure of recombinant 13C,15N-labeled oryctin by heteronuclear NMR spectroscopy. Oryctin exhibits a fold similar to that of Kazal-type serine protease inhibitors but has a unique additional C-terminal α-helix. We performed protease inhibition assays of oryctin against several bacterial and eukaryotic proteases. Oryctin does inhibit the following serine proteases: α-chymotrypsin, endopeptidase K, subtilisin Carlsberg, and leukocyte elastase, with Ki values of 3.9 × 10−10 m, 6.2 × 10−10 m, 1.4 × 10−9 m, and 1.2 × 10−8 m, respectively. Although the target molecule of oryctin in the beetle hemolymph remains obscure, our results showed that oryctin is a novel single domain Kazal-type inhibitor and could play a key role in protecting against bacterial infections. PMID:20630859

Radioactive cDNA microarray in neurospsychiatry

International Nuclear Information System (INIS)

Choe, Jae Gol; Shin, Kyung Ho; Lee, Min Soo; Kim, Meyoung Kon

2003-01-01

Microarray technology allows the simultaneous analysis of gene expression patterns of thousands of genes, in a systematic fashion, under a similar set of experimental conditions, thus making the data highly comparable. In some cases arrays are used simply as a primary screen leading to downstream molecular characterization of individual gene candidates. In other cases, the goal of expression profiling is to begin to identify complex regulatory networks underlying developmental processes and disease states. Microarrays were originally used with cell lines or other simple model systems. More recently, microarrays have been used in the analysis of more complex biological tissues including neural systems and the brain. The application of cDNA arrays in neuropsychiatry has lagged behind other fields for a number of reasons. These include a requirement for a large amount of input probe RNA in fluorescent-glass based array systems and the cellular complexity introduced by multicellular brain and neural tissues. An additional factor that impacts the general use of microarrays in neuropsychiatry is the lack of availability of sequenced clone sets from model systems. While human cDNA clones have been widely available, high quality rat, mouse, and drosophilae, among others are just becoming widely available. A final factor in the application of cDNA microarrays in neuropsychiatry is cost of commercial arrays. As academic microarray facilitates become more commonplace custom made arrays will become more widely available at a lower cost allowing more widespread applications. In summary, microarray technology is rapidly having an impact on many areas of biomedical research. Radioisotope-nylon based microarrays offer alternatives that may in some cases be more sensitive, flexible, inexpensive, and universal as compared to other array formats, such as fluorescent-glass arrays. In some situations of limited RNA or exotic species, radioactive membrane microarrays may be the most

Radioactive cDNA microarray in neurospsychiatry

Energy Technology Data Exchange (ETDEWEB)

Choe, Jae Gol; Shin, Kyung Ho; Lee, Min Soo; Kim, Meyoung Kon [Korea University Medical School, Seoul (Korea, Republic of)

2003-02-01

Microarray technology allows the simultaneous analysis of gene expression patterns of thousands of genes, in a systematic fashion, under a similar set of experimental conditions, thus making the data highly comparable. In some cases arrays are used simply as a primary screen leading to downstream molecular characterization of individual gene candidates. In other cases, the goal of expression profiling is to begin to identify complex regulatory networks underlying developmental processes and disease states. Microarrays were originally used with cell lines or other simple model systems. More recently, microarrays have been used in the analysis of more complex biological tissues including neural systems and the brain. The application of cDNA arrays in neuropsychiatry has lagged behind other fields for a number of reasons. These include a requirement for a large amount of input probe RNA in fluorescent-glass based array systems and the cellular complexity introduced by multicellular brain and neural tissues. An additional factor that impacts the general use of microarrays in neuropsychiatry is the lack of availability of sequenced clone sets from model systems. While human cDNA clones have been widely available, high quality rat, mouse, and drosophilae, among others are just becoming widely available. A final factor in the application of cDNA microarrays in neuropsychiatry is cost of commercial arrays. As academic microarray facilitates become more commonplace custom made arrays will become more widely available at a lower cost allowing more widespread applications. In summary, microarray technology is rapidly having an impact on many areas of biomedical research. Radioisotope-nylon based microarrays offer alternatives that may in some cases be more sensitive, flexible, inexpensive, and universal as compared to other array formats, such as fluorescent-glass arrays. In some situations of limited RNA or exotic species, radioactive membrane microarrays may be the most

Spatial Distribution of Adult Anthonomus grandis Boheman (Coleoptera: Curculionidae) and Damage to Cotton Flower Buds Due to Feeding and Oviposition.

Science.gov (United States)

Grigolli, J F J; Souza, L A; Fernandes, M G; Busoli, A C

2017-08-01

The cotton boll weevil Anthonomus grandis Boheman (Coleoptera: Curculionidae) is the main pest in cotton crop around the world, directly affecting cotton production. In order to establish a sequential sampling plan, it is crucial to understand the spatial distribution of the pest population and the damage it causes to the crop through the different developmental stages of cotton plants. Therefore, this study aimed to investigate the spatial distribution of adults in the cultivation area and their oviposition and feeding behavior throughout the development of the cotton plants. The experiment was conducted in Maracaju, Mato Grosso do Sul, Brazil, in the 2012/2013 and 2013/2014 growing seasons, in an area of 10,000 m 2 , planted with the cotton cultivar FM 993. The experimental area was divided into 100 plots of 100 m 2 (10 × 10 m) each, and five plants per plot were sampled weekly throughout the crop cycle. The number of flower buds with feeding and oviposition punctures and of adult A. grandis was recorded throughout the crop cycle in five plants per plot. After determining the aggregation indices (variance/mean ratio, Morisita's index, exponent k of the negative binomial distribution, and Green's coefficient) and adjusting the frequencies observed in the field to the distribution of frequencies (Poisson, negative binomial, and positive binomial) using the chi-squared test, it was observed that flower buds with punctures derived from feeding, oviposition, and feeding + oviposition showed an aggregated distribution in the cultivation area until 85 days after emergence and a random distribution after this stage. The adults of A. grandis presented a random distribution in the cultivation area.

Effect of moisture availability on wood density and vessel characteristics of Eucalyptus grandis in the warm temperate region of South Africa

CSIR Research Space (South Africa)

Naidoo, Sasha

2006-09-01

Full Text Available intolerant of adverse conditions, and performs poorly when planted on shallow soils and/or on dry sites. A study was conducted to assess the effect of moisture availability on the wood density and vessel characteristics of E. grandis grown in the warm...

A BIOINFORMATIC STRATEGY TO RAPIDLY CHARACTERIZE CDNA LIBRARIES

Science.gov (United States)

A Bioinformatic Strategy to Rapidly Characterize cDNA LibrariesG. Charles Ostermeier1, David J. Dix2 and Stephen A. Krawetz1.1Departments of Obstetrics and Gynecology, Center for Molecular Medicine and Genetics, & Institute for Scientific Computing, Wayne State Univer...

Within plant distribution of Anthonomus grandis (Coleoptera: Curculionidae feeding and oviposition damages in cotton cultivars Distribuição vertical de botões florais com danos de alimentação e de oviposição de Anthonomus grandis (Coleoptera: Curculionidae em cultivares de algodoeiro

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José Fernando Jurca Grigolli

2013-02-01

Full Text Available The feeding and oviposition behavior of boll weevil in new cotton cultivars is essential for an adequate management. The objective of this study was to evaluate the vertical distribution of squares punctured for feeding and oviposition of the pest in the cultivars NuOPAL, DeltaOPAL, FMT-701, FMX-910 and FMX-993, and record the most and least preferred times of feeding and oviposition. The number of squares used for boll weevil feeding and oviposition were evaluated weekly in three parts of plant canopy. It was observed that, regardless the cultivar, A. grandis preferred to lay eggs in squares located in the upper part and feed on squares in the middle and upper parts. The boll weevil preferred to feed on cultivar FMT-701 in the beginning of the period of cotton flowering and fruiting, and the cultivars NuOPAL, DeltaOPAL, FMX-910 and FMX-993 throughout the whole period of flowering and fruiting. A. grandis preferred to lay eggs on cultivars NuOPAL, FMT-701 and FMX-993 at the beginning and end of flowering and fruiting of plants, while the cultivars DeltaOPAL and FMX-910 are used for oviposition throughout the period of flowering and fruiting.O conhecimento do comportamento de alimentação e de oviposição de Anthonomus grandis em cultivares recentes de algodoeiro é essencial para seu manejo. Neste trabalho, objetivou-se avaliar a distribuição vertical de botões florais com orifícios de alimentação e de oviposição da praga nas cultivares NuOPAL, DeltaOPAL, FMT-701, FMX-910 e FMX-993, bem como registrar as épocas de maior e menor preferência alimentar e de oviposição. O experimento foi conduzido em Jaboticabal, SP, Brasil, safra 2010/2011. Foram realizadas avaliações semanais, baseadas no número de botões florais, utilizados para alimentação e para oviposição pelo bicudo-do-algodoeiro, em três regiões do dossel das plantas. Observou-se que A. grandis preferiu ovipositar em botões florais localizados no terço superior das

[Cloning and sequencing of KIR2DL1 framework gene cDNA and identification of a novel allele].

Science.gov (United States)

Sun, Ge; Wang, Chang; Zhen, Jianxin; Zhang, Guobin; Xu, Yunping; Deng, Zhihui

2016-10-01

To develop an assay for cDNA cloning and haplotype sequencing of KIR2DL1 framework gene and determine the genotype of an ethnic Han from southern China. Total RNA was isolated from peripheral blood sample, and complementary DNA (cDNA) transcript was synthesized by RT-PCR. The entire coding sequence of the KIR2DL1 framework gene was amplified with a pair of KIR2DL1-specific PCR primers. The PCR products with a length of approximately 1.2 kb were then subjected to cloning and haplotype sequencing. A specific target fragment of the KIR2DL1 framework gene was obtained. Following allele separation, a wild-type KIR2DL1*00302 allele and a novel variant allele, KIR2DL1*031, were identified. Sequence alignment with KIR2DL1 alleles from the IPD-KIR Database showed that the novel allele KIR2DL1*031 has differed from the closest allele KIR2DL1*00302 by a non-synonymous mutation at CDS nt 188A>G (codon 42 GAG>GGG) in exon 4, which has caused an amino acid change Glu42Gly. The sequence of the novel allele KIR2DL1*031 was submitted to GenBank under the accession number KP025960 and to the IPD-KIR Database under the submission number IWS40001982. A name KIR2DL1*031 has been officially assigned by the World Health Organization (WHO) Nomenclature Committee. An assay for cDNA cloning and haplotype sequencing of KIR2DL1 has been established, which has a broad applications in KIR studies at allelic level.

Production of a full-length infectious GFP-tagged cDNA clone of Beet mild yellowing virus for the study of plant-polerovirus interactions.

Science.gov (United States)

Stevens, Mark; Viganó, Felicita

2007-04-01

The full-length cDNA of Beet mild yellowing virus (Broom's Barn isolate) was sequenced and cloned into the vector pLitmus 29 (pBMYV-BBfl). The sequence of BMYV-BBfl (5721 bases) shared 96% and 98% nucleotide identity with the other complete sequences of BMYV (BMYV-2ITB, France and BMYV-IPP, Germany respectively). Full-length capped RNA transcripts of pBMYV-BBfl were synthesised and found to be biologically active in Arabidopsis thaliana protoplasts following electroporation or PEG inoculation when the protoplasts were subsequently analysed using serological and molecular methods. The BMYV sequence was modified by inserting DNA that encoded the jellyfish green fluorescent protein (GFP) into the P5 gene close to its 3' end. A. thaliana protoplasts electroporated with these RNA transcripts were biologically active and up to 2% of transfected protoplasts showed GFP-specific fluorescence. The exploitation of these cDNA clones for the study of the biology of beet poleroviruses is discussed.

Transcriptomic identification of candidate genes involved in sunflower responses to chilling and salt stresses based on cDNA microarray analysis

Directory of Open Access Journals (Sweden)

Paniego Norma

2008-01-01

Full Text Available Abstract Background Considering that sunflower production is expanding to arid regions, tolerance to abiotic stresses as drought, low temperatures and salinity arises as one of the main constrains nowadays. Differential organ-specific sunflower ESTs (expressed sequence tags were previously generated by a subtractive hybridization method that included a considerable number of putative abiotic stress associated sequences. The objective of this work is to analyze concerted gene expression profiles of organ-specific ESTs by fluorescence microarray assay, in response to high sodium chloride concentration and chilling treatments with the aim to identify and follow up candidate genes for early responses to abiotic stress in sunflower. Results Abiotic-related expressed genes were the target of this characterization through a gene expression analysis using an organ-specific cDNA fluorescence microarray approach in response to high salinity and low temperatures. The experiment included three independent replicates from leaf samples. We analyzed 317 unigenes previously isolated from differential organ-specific cDNA libraries from leaf, stem and flower at R1 and R4 developmental stage. A statistical analysis based on mean comparison by ANOVA and ordination by Principal Component Analysis allowed the detection of 80 candidate genes for either salinity and/or chilling stresses. Out of them, 50 genes were up or down regulated under both stresses, supporting common regulatory mechanisms and general responses to chilling and salinity. Interestingly 15 and 12 sequences were up regulated or down regulated specifically in one stress but not in the other, respectively. These genes are potentially involved in different regulatory mechanisms including transcription/translation/protein degradation/protein folding/ROS production or ROS-scavenging. Differential gene expression patterns were confirmed by qRT-PCR for 12.5% of the microarray candidate sequences. Conclusion

A technique to identify annual growth rings in Eucalyptus grandis using annual measurements of diameter at breast height and gamma ray densitometry

CSIR Research Space (South Africa)

Naidoo, Sasha

2010-06-01

Full Text Available A technique was developed to identify annual growth rings in E. grandis using a combination of annual measurements of diameter at breast height (DBH) from permanent sample plot (PSP) datasets and bark-pith density profiles. By assessing the pattern...

Cloning and expression of a cDNA encoding human sterol carrier protein 2

International Nuclear Information System (INIS)

Yamamoto, Ritsu; Kallen, C.B.; Babalola, G.O.; Rennert, H.; Strauss, J.F. III; Billheimer, J.T.

1991-01-01

The authors report the cloning and expression of a cDNA encoding human sterol carrier protein 2 (SCP 2 ). The 1.3-kilobase (kb) cDNA contains an open reading frame which encompasses a 143-amino acid sequence which is 89% identical to the rat SCP 2 amino acid sequence. The deduced amino acid sequence of the polypeptide reveals a 20-residue amino-terminal leader sequence in front of the mature polypeptide, which contains a carboxyl-terminal tripeptide (Ala-Lys-Leu) related to the peroxisome targeting sequence. The expressed cDNA in COS-7 cells yields a 15.3-kDa polypeptide and increased amounts of a 13.2-kDa polypeptide, both reacting with a specific rabbit antiserum to rat liver SCP 2 . The cDNA insert hybridizes with 3.2- and 1.8-kb mRNA species in human liver poly(A) + RNA. In human fibroblasts and placenta the 1.8-kb mRNA was most abundant. Southern blot analysis suggests either that there are multiple copies of the SCP 2 gene in the human genome or that the SCP 2 gene is very large. Coexpression of the SCP 2 cDNA with expression vectors for cholesterol side-chain cleavage enzyme and adrenodoxin resulted in a 2.5-fold enhancement of progestin synthesis over that obtained with expression of the steroidogenic enzyme system alone. These findings are concordant with the notion that SCP 2 plays a role in regulating steroidogenesis, among other possible functions

Molecular cloning of a cDNA encoding the precursor of adenoregulin from frog skin. Relationships with the vertebrate defensive peptides, dermaseptins.

Science.gov (United States)

Amiche, M; Ducancel, F; Lajeunesse, E; Boulain, J C; Ménez, A; Nicolas, P

1993-03-31

Adenoregulin has recently been isolated from Phyllomedusa skin as a 33 amino acid residues peptide which enhanced binding of agonists to the A1 adenosine receptor. In order to study the structure of the precursor of adenoregulin we constructed a cDNA library from mRNAs extracted from the skin of Phyllomedusa bicolor. We detected the complete nucleotide sequence of a cDNA encoding the adenoregulin biosynthetic precursor. The deduced sequence of the precursor is 81 amino acids long, exhibits a putative signal sequence at the NH2 terminus and contains a single copy of the biologically active peptide at the COOH terminus. Structural and conformational homologies that are observed between adenoregulin and the dermaseptins, antimicrobial peptides exhibiting strong membranolytic activities against various pathogenic agents, suggest that adenoregulin is an additional member of the growing family of cytotropic antimicrobial peptides that allow vertebrate animals to defend themselves against microorganisms. As such, the adenosine receptor regulating activity of adenoregulin could be due to its ability to interact with and disrupt membranes lipid bilayers.

Construction of a cDNA microarray derived from the ascidian Ciona intestinalis.

Science.gov (United States)

Azumi, Kaoru; Takahashi, Hiroki; Miki, Yasufumi; Fujie, Manabu; Usami, Takeshi; Ishikawa, Hisayoshi; Kitayama, Atsusi; Satou, Yutaka; Ueno, Naoto; Satoh, Nori

2003-10-01

A cDNA microarray was constructed from a basal chordate, the ascidian Ciona intestinalis. The draft genome of Ciona has been read and inferred to contain approximately 16,000 protein-coding genes, and cDNAs for transcripts of 13,464 genes have been characterized and compiled as the "Ciona intestinalis Gene Collection Release I". In the present study, we constructed a cDNA microarray of these 13,464 Ciona genes. A preliminary experiment with Cy3- and Cy5-labeled probes showed extensive differential gene expression between fertilized eggs and larvae. In addition, there was a good correlation between results obtained by the present microarray analysis and those from previous EST analyses. This first microarray of a large collection of Ciona intestinalis cDNA clones should facilitate the analysis of global gene expression and gene networks during the embryogenesis of basal chordates.

ESTABELECIMENTO A CAMPO DE MUDAS DE Eucalyptus grandis MICORRIZADAS COM Pisolithus microcarpus (UFSC Pt 116 EM SOLO ARENOSO

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Andréa Hentz de Mello

2009-01-01

Full Text Available The aim of this work was to evaluate the survival and the initial growth of mycorrhizated eucalypts with Pisolithus microcarpus (UFSC Pt 116 ectomycorrhizal fungus, after its transplant to area subject to the arenization process in São Francisco de Assis, RS. The area was divided into four blocks, each one with four treatments (fertile turf with and without mycorrhizae, Quartzarenic Neosoil with and without mycorrhizae. Each parcel was composed of 16 seedlings arranged in four lines in the spacing of 1,5 m x 1,5 m, totalizing in each block 64 seedlings. 90 days after the planting in the field, the eucaliptus seedlings produced in turf with fungus in the fertile substratum presented a survival rate of 100 %, whereas for those produced in fertile turf without fingi, the survival rate was 92 %. The seedlings produced in the Quartzarenic Neosoil with and without mycorrhizae had a survival rate varying around 98 and 89 %, respectively. The produced seedlings with turf and fungus showed significant differences in height and stem diameter. This study showed that the Eucalyptus grandis seedlings produced in substratum fertile turf and inoculated with the Pisolithus microcarpus (UFSC Pt 116 isolated may maintain good development and establishment in the field.

An enzyme-immunobinding assay for fast screening of expression of tissue plasminogen activator cDNA in E. coli

International Nuclear Information System (INIS)

Tang, J.C.T.; Li, S.H.

1984-01-01

Tissue plasminogen activator (TPA) has been isolated from normal human tissues and certain human cell lines in culture. The enzyme is a serine protease which converts an inactive zymogen, plasminogen to plasmin, and causes lysis of fibrin clots. The high affinity of TPA for fibrin indicates that it is a potential thrombolytic agent and is superior to urokinase-like plasminogen activators. Recently, TPA has been cloned and expressed in E. coli. Using TPA as a model protein, the authors report here the development of a direct, sensitive enzyme-immunoassay for the screening of a cDNA expression library using specific antibodies and peroxidase-labeled second antibody

Proteômica quantitativa e metabolômica do híbrido Eucalyptus grandis x E. camaldulensis, tolerante e susceptível ao déficit hídrico

OpenAIRE

Janaina de Santana Borges

2016-01-01

O E. grandis x E. camaldulensis possui características favoráveis de adaptação à seca, conferidas pelo E. camaldulensis e qualidade da madeira para papel e celulose, conferida pelo E. grandis. Esta adaptação à seca está relacionada a fatores fisiológicos e também moleculares, expressos em sua proteoma e metaboloma, que se alteram na presença do estresse. Objetiva-se neste trabalho estudar as respostas fisiológicas, proteômicas e metabolômicas (metabólitos primários) diferencialmente expressos...

RFLP for Duchenne muscular dystrophy cDNA clone 44-1

Energy Technology Data Exchange (ETDEWEB)

Laing, N G; Siddique, T; Bartlett, R J; Yamaoka, L H; Chen, J C; Walker, A P; Hung, W Y; Roses, A D [Duke Univ. Medical Center, Durham, NC (USA)

1988-07-25

Clone 44-1 is one of six cDNA clones which comprise the cDNA for the Duchenne muscular dystrophy gene. It is a 0.9kb fragment in the EcoR1 site of Bluescript. Taq1 (TlCGA) identifies two alleles with bands at 6.8 and 5.7kb, as well as four constant bands at 4.8, 3.9, 3.5 and 2.5kb. Its frequency was studied in 62 unrelated individuals. Mendelian inheritance was demonstrated in one three generation and three two generation informative families, 26 individuals. There were no problems on RFLP analysis under normal stringency conditions.

Amenizante Orgânico e Eucalyptus grandis para Fitoestabilização de Solo Contaminado com Cobre

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Rudinei De Marco

2017-04-01

Full Text Available RESUMO Este trabalho avaliou a utilização de turfa como amenizante e Eucalyptus grandis como técnica de fitoestabilização para remediar solo contaminado com cobre. O trabalho foi conduzido sob delineamento inteiramente casualizado em arranjo fatorial (2 × 6, sendo sem e com adição de turfa (200 mL L-1 de solo e seis doses de cobre adicionadas ao solo (0 mg kg-1, 60 mg kg-1, 120 mg kg-1, 180 mg kg-1, 240 mg kg-1, 300 mg kg-1 de solo, com seis repetições. Avaliaram-se a altura das mudas, o diâmetro do colo, a massa seca radicular e aérea, a área superficial específica, os teores e a quantidade acumulada de cobre no sistema radicular e na parte aérea e o índice de translocação. A adição de turfa possibilitou efeito amenizante da contaminação por cobre e favoreceu o crescimento das mudas. As mudas de Eucalyptus grandis apresentam elevado acúmulo de cobre nas raízes, o que permite indicá-la como promissora para fins de fitoestabilização de solos contaminados com cobre.

Genome-Wide Characterization and Expression Profiling of the AUXIN RESPONSE FACTOR (ARF) Gene Family in Eucalyptus grandis

Science.gov (United States)

Yu, Hong; Soler, Marçal; Mila, Isabelle; San Clemente, Hélène; Savelli, Bruno; Dunand, Christophe; Paiva, Jorge A. P.; Myburg, Alexander A.; Bouzayen, Mondher; Grima-Pettenati, Jacqueline; Cassan-Wang, Hua

2014-01-01

Auxin is a central hormone involved in a wide range of developmental processes including the specification of vascular stem cells. Auxin Response Factors (ARF) are important actors of the auxin signalling pathway, regulating the transcription of auxin-responsive genes through direct binding to their promoters. The recent availability of the Eucalyptus grandis genome sequence allowed us to examine the characteristics and evolutionary history of this gene family in a woody plant of high economic importance. With 17 members, the E. grandis ARF gene family is slightly contracted, as compared to those of most angiosperms studied hitherto, lacking traces of duplication events. In silico analysis of alternative transcripts and gene truncation suggested that these two mechanisms were preeminent in shaping the functional diversity of the ARF family in Eucalyptus. Comparative phylogenetic analyses with genomes of other taxonomic lineages revealed the presence of a new ARF clade found preferentially in woody and/or perennial plants. High-throughput expression profiling among different organs and tissues and in response to environmental cues highlighted genes expressed in vascular cambium and/or developing xylem, responding dynamically to various environmental stimuli. Finally, this study allowed identification of three ARF candidates potentially involved in the auxin-regulated transcriptional program underlying wood formation. PMID:25269088

Genome-wide characterization and expression profiling of the AUXIN RESPONSE FACTOR (ARF gene family in Eucalyptus grandis.

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Hong Yu

Full Text Available Auxin is a central hormone involved in a wide range of developmental processes including the specification of vascular stem cells. Auxin Response Factors (ARF are important actors of the auxin signalling pathway, regulating the transcription of auxin-responsive genes through direct binding to their promoters. The recent availability of the Eucalyptus grandis genome sequence allowed us to examine the characteristics and evolutionary history of this gene family in a woody plant of high economic importance. With 17 members, the E. grandis ARF gene family is slightly contracted, as compared to those of most angiosperms studied hitherto, lacking traces of duplication events. In silico analysis of alternative transcripts and gene truncation suggested that these two mechanisms were preeminent in shaping the functional diversity of the ARF family in Eucalyptus. Comparative phylogenetic analyses with genomes of other taxonomic lineages revealed the presence of a new ARF clade found preferentially in woody and/or perennial plants. High-throughput expression profiling among different organs and tissues and in response to environmental cues highlighted genes expressed in vascular cambium and/or developing xylem, responding dynamically to various environmental stimuli. Finally, this study allowed identification of three ARF candidates potentially involved in the auxin-regulated transcriptional program underlying wood formation.

Cloning, molecular characterization and expression of a cDNA encoding a functional NADH-cytochrome b5 reductase from Mucor racemosus PTCC 5305 in E. coli

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NED A SETAYESH

2009-01-01

Full Text Available The present work aims to study a new NADH-cytochrome b5 reductase (cb5r from Mucor racemosus PTCC 5305. A cDNA coding for cb s r was isolated from a Mucor racemosus PTCC 5305 cDNA library. The nucleotide sequence of the cDNA including coding and sequences flanking regions was determined. The open reading frame starting from ATG and ending with TAG stop codon encoded 228 amino acids and displayed the closest similarity (73% with Mortierella alpina cb s r. Lack of hydrophobic residues in the N-terminal sequence was apparent, suggesting that the enzyme is a soluble isoform. The coding sequence was then cloned in the pET16b transcription vector carrying an N-terminal-linked His-Tag® sequence and expressed in Escherichia coli BL21 (DE3. The enzyme was then homogeneously purified by a metal affinity column. The recombinant Mucor enzyme was shown to have its optimal activity at pH and temperature of about 7.5 and 40 °C, respectively. The apparent Km value was calculated to be 13 μM for ferricyanide. To our knowledge, this is the first report on cloning and expression of a native fungal soluble isoform of NADH-cytochrome b5 reductase in E. coli.

Avoiding cross hybridization by choosing nonredundant targets on cDNA arrays

DEFF Research Database (Denmark)

Nielsen, Henrik Bjørn; Knudsen, Steen

2002-01-01

PROBEWIZ designs PCR primers for amplifying probes for cDNA arrays. The probes are designed to have minimal homology to other expressed sequences from a given organism. The primer selection is based on user-defined penalties for homology, primer quality, and proximity to the 3' end.......PROBEWIZ designs PCR primers for amplifying probes for cDNA arrays. The probes are designed to have minimal homology to other expressed sequences from a given organism. The primer selection is based on user-defined penalties for homology, primer quality, and proximity to the 3' end....

cDNA cloning and mRNA expression of heat shock protein 70 gene ...

African Journals Online (AJOL)

In this study, the full-length heat shock protein 70 of Tegillarca granosa was cloned from cDNA library by rapid amplification of cDNA end (RACE). The open reading frame (ORF) of heat shock protein 70 was 1968 bp, and it encoded a protein of 655 amino acids with a predicted molecular weight of 71.48 kDa and an ...

Analysis of Cry8Ka5-binding proteins from Anthonomus grandis (Coleoptera: Curculionidae) midgut.

Science.gov (United States)

Nakasu, Erich Y T; Firmino, Alexandre A P; Dias, Simoni C; Rocha, Thales L; Ramos, Hudson B; Oliveira, Gustavo R; Lucena, Wagner; Carlini, Célia R; Grossi-de-Sá, Maria Fátima

2010-07-01

Biotech crops expressing Bacillus thuringiensis Cry toxins present a valuable approach for insect control. Cry8Ka5, which is highly toxic to the cotton boll weevil (Anthonomus grandis), was used as a model to study toxin-ligand interactions. Three Cry-binding proteins were detected after toxin overlay assays. Following de novo sequencing, a heat-shock cognate protein and a V-ATPase were identified, whilst a approximately 120 kDa protein remained unknown. Additional Cry8Ka5-binding proteins were visualized by two-dimensional gel electrophoresis ligand blots. (c) 2010 Elsevier Inc. All rights reserved.

Identification of cDNA clones expressing immunodiagnostic antigens from Trichinella spiralis

International Nuclear Information System (INIS)

Zarlenga, D.; Gamble, H.R.

1987-01-01

A cDNA expression library was built in lambda gt11 phage using poly A mRNA isolated from Trichinella spiralis muscle stage larvae. This library was screened with rabbit antibodies to parasite excretory-secretory (ES) products and greater than 180 clones were isolated. Thirteen clones producing highly immunogenic protein antigens were plaque purified and rescreened with pig antisera to T.spiralis, Trichuris suis or Ascaris suum to identify clones producing epitopes specific to T.spiralis ES products, only. Two clones, TsAc-2 and TsAc-8, which displayed strong interactions with pig antisera to T. spiralis were lysogenized in E. coli Y1089 and the protein extracted. Western blots of the crude fusion proteins revealed molecular weights of 133 kD and 129 kD, respectively. Northern blot analysis of total RNA with 32 P labelled cDNA:lambda gt11 probes indicated single RNA transcripts for each clone with molecular sizes corresponding to 800-850 nucleotides. dscDNA inserts were estimated by southern blot analysis to be 500 bp and 340 bp, respectively, with no cross-hybridization observed between the cloned sequences. Dot blots using pig sera to screen crude fusion protein preparations, total bacterial protein (negative controls) and crude worm extract or ES products from T.spiralis, T.suis and A.suum (positive controls) corroborated the specificity and sensitivity of these clones as potential diagnostic antigens for swine trichinellosis

Cloning, sequencing and expression of cDNA encoding growth ...

Indian Academy of Sciences (India)

Unknown

of medicine, animal husbandry, fish farming and animal ..... northern pike (Esox lucius) growth hormone; Mol. Mar. Biol. ... prolactin 1-luciferase fusion gene in African catfish and ... 1988 Cloning and sequencing of cDNA that encodes goat.

SPECTRORADIOMETRY IN THE VISIBLE AND NEAR INFRARED REGION ON A STAND OF Eucalyptus grandis Hill ex-Maiden

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Catize Brandelero

2012-03-01

Full Text Available http://dx.doi.org/10.5902/198050985093Reflectance readings in border and inner tree leaves in a Eucalyptus grandis stand, in São Pedro das Missões, Rio Grande do Sul state, were analyzed in the regions of the visible electromagnetic spectrum and the nearby infrared, by using spectrum radiometry. The area was divided in two parts: border and center stands. In order to collect the material, the crown was divided in three parts (superior, medium and inferior, so that it would be possible to differentiate the positions of leaf collections in each area. Three trees were sampled in each area, adding up to six trees, for each tree, 60 isolated leaves were collected, 20 in each position. The reflectance readings were carried out through FieldSpec®3 spectrum radiometer and the final results were segmented in the visible and nearby infrared spectral bands. The statistical analysis was made on the basis of several tests, among them Tukey HSD test, in order to compare the averages of the visible region, which, according to ANOVA, present significant differences. It is concluded that the collecting indicating class of leaves for the spectrum radiometric analysis in the visible region are preferably the 5 one (tree in the center, reading in the medium part and #3 one (border tree, reading superior part.

Analysis of a cDNA clone expressing a human autoimmune antigen: full-length sequence of the U2 small nuclear RNA-associated B antigen

International Nuclear Information System (INIS)

Habets, W.J.; Sillekens, P.T.G.; Hoet, M.H.; Schalken, J.A.; Roebroek, A.J.M.; Leunissen, J.A.M.; Van de Ven, W.J.M.; Van Venrooij, W.J.

1987-01-01

A U2 small nuclear RNA-associated protein, designated B'', was recently identified as the target antigen for autoimmune sera from certain patients with systemic lupus erythematosus and other rheumatic diseases. Such antibodies enabled them to isolate cDNA clone λHB''-1 from a phage λgt11 expression library. This clone appeared to code for the B'' protein as established by in vitro translation of hybrid-selected mRNA. The identity of clone λHB''-1 was further confirmed by partial peptide mapping and analysis of the reactivity of the recombinant antigen with monospecific and monoclonal antibodies. Analysis of the nucleotide sequence of the 1015-base-pair cDNA insert of clone λHB''-1 revealed a large open reading frame of 800 nucleotides containing the coding sequence for a polypeptide of 25,457 daltons. In vitro transcription of the λHB''-1 cDNA insert and subsequent translation resulted in a protein product with the molecular size of the B'' protein. These data demonstrate that clone λHB''-1 contains the complete coding sequence of this antigen. The deduced polypeptide sequence contains three very hydrophilic regions that might constitute RNA binding sites and/or antigenic determinants. These findings might have implications both for the understanding of the pathogenesis of rheumatic diseases as well as for the elucidation of the biological function of autoimmune antigens

Isolation of a kernel oleoyl-ACP thioesterase gene from the oil palm ...

African Journals Online (AJOL)

We have isolated a cDNA clone from the developing kernel of the oil palm Elaeis guineensis which encodes a thioesterase enzyme. Its highest homology was to the Brassica napus oleoyl-ACP thioesterase with which it had 72% homology at the nucleotide level, over the coding region examined, and 83% identity (90% ...

CARACTERÍSTICAS TECNOLÓGICAS DA MADEIRA DE ÁRVORES MATRIZES DE Eucalyptus grandis

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Denis L. G. Fernandes

2004-06-01

Full Text Available O objetivo do presente trabalho foi caracterizar árvores matrizes de Eucalyptus grandis em relação a características tecnológicas da madeira. Foram selecionadas 63 árvores matrizes, pertencentes a um povoamento comercial localizado no litoral norte do Rio Grande do Sul. As árvores foram abatidas e, com base nos dados dendrométricos, calculou-se o volume comercial com e sem casca, o fator de forma, a conicidade e a relação altura/diâmetro. Quanto à madeira, foi analisada a massa específica básica, os percentuais volumétricos de cerne, alburno e casca, as rachaduras de topo das toras e das tábuas, os empenamentos e os defeitos visuais das tábuas, tais como nós e bolsa de resina. A massa específica básica, cuja média inclui a madeira dessa espécie entre as moderadamente leves a pesadas, mostrou uma tendência decrescente entre o DAP e 25% da altura comercial e, a partir daí, crescente até 100% da mesma. O percentual volumétrico de cerne apresentou valores médios crescentes desde a base até 25% da altura comercial e, a partir daí, diminuiu até 100% da altura comercial, sendo que a média geral foi de 75,7%. O comprimento médio das rachaduras de topo em tábuas de Eucalyptus grandis foi de 63%. A alta porcentagem de rachaduras de topo foi atribuída à posição de retirada das tábuas, próxima à medula. Já a variabilidade das rachaduras de topo encontrada para as toras (CV=60% possibilita a seleção de árvores com menor tendência em apresentar esse defeito.

René Descartes, the Robots and the Soul in the debate between Tommaso Ceva and Guido Grandi

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Alfredo Serrai

2016-05-01

Full Text Available Descartes outlined human intelligence as a divine gift, albeit in a system governed by universal laws of physics and mathematics. In the eighteenth century the camaldolite Guido Grandi and Tommaso Ceva, both mathematicians, had a dispute about the interpretation of the Cartesian model of human intelligence. This case provides an opportunity to retrace the path of a speculative techno-philosophical question concerning the human possibility to equipping robots with the ability to experience a self-consciousness.

cDNA sequence and tissue distribution of the mRNA for bovine and murine p11, the S100-related light chain of the protein-tyrosine kinase substrate p36 (calpactin I)

DEFF Research Database (Denmark)

Saris, Chris J M; Kristensen, Torsten; D’Eustachio, Peter

1987-01-01

We have isolated and sequenced cDNA clones of bovine nd murine pl 1 mRNAs. The nonpolyadenylated mRNAs are predicted to be 614 and 600 nucleotides, respectively. The p l l mRNAs both contain a 291 nucleotide open reading frame, preceded by a 5”untranslated region of 73 nucleotides in bovine p l l m...

Replication of Chilo iridescent virus in the cotton boll weevil, Anthonomus grandis, and development of an infectivity assay.

Science.gov (United States)

Henderson, C W; Johnson, C L; Lodhi, S A; Bilimoria, S L

2001-01-01

The boll weevil, Anthonomus grandis Boheman, is a devastating pest of cotton. Chemical pesticides are problematic due to relative lack of target specificity and resistance. Microbial pesticides may provide viable alternatives because of their narrow host range. Chilo iridescent virus (CIV) is the type species for genus Iridovirus, family Iridoviridae: large, icosahedral cytoplasmic viruses containing a double-stranded DNA genome. Earlier work suggested that CIV replicated in the boll weevil; however, efficiency or production of infectious virus was not established. We showed that CIV undergoes a productive cycle in A. grandis. CIV DNA levels in boll weevil pupae increased significantly from 0 to 3 days post infection. Moreover, virogenic stromata and complete virus particles were observed in the cytoplasm by 7 days. An endpoint dilution assay using viral DNA replication as indicator suggested a 10(5)-fold increase in infectious virus titer over 7 days. This is the first such demonstration in larval infections with genus Iridovirus. Our study establishes that CIV undergoes a productive cycle in the boll weevil and provides an important and useful model system for replication at the organismal level. These results have important implications for the potential of CIV and its components in boll weevil control.

Cloning and expression of a cDNA coding for a human monocyte-derived plasminogen activator inhibitor.

OpenAIRE

Antalis, T M; Clark, M A; Barnes, T; Lehrbach, P R; Devine, P L; Schevzov, G; Goss, N H; Stephens, R W; Tolstoshev, P

1988-01-01

Human monocyte-derived plasminogen activator inhibitor (mPAI-2) was purified to homogeneity from the U937 cell line and partially sequenced. Oligonucleotide probes derived from this sequence were used to screen a cDNA library prepared from U937 cells. One positive clone was sequenced and contained most of the coding sequence as well as a long incomplete 3' untranslated region (1112 base pairs). This cDNA sequence was shown to encode mPAI-2 by hybrid-select translation. A cDNA clone encoding t...

Th, Pa and U isotopes in an echinoderm, Encope grandis

International Nuclear Information System (INIS)

Omura, Akio; Ku, Teh-Lung.

1979-01-01

The application of 230 Th and 231 Pa growth methods to the hard tissues of living things, which are effective for the radiometric age measurement for latter Quaternary period, has been limited to certain corals, therefore it has been scarcely utilized in other areas than coral reefs. Reef coral fossils (Porites) were obtained from terrace deposits of Magdalena Island in Southern Baja California, and the methods were applied to them. At the time, the isotope compositions of Th, Pa and U in the shells of echinoderm Encope Grandis and of the living samples were examined. The estimated ages were in agreement with those of coral. It suggests that the reliable 230 Th and 231 Pa ages of sea-urchin fossils were presented for the first time and that the method is applicable to such fossils only if the conditions can be met. The results are highly significant, since the method may be used in other areas than coral reefs. (J.P.N.)

Relative contributions of copper oxide nanoparticles and dissolved copper to Cu uptake kinetics of Gulf killifish (Fundulus grandis) embryos

Science.gov (United States)

Jiang, Chuanjia; Castellon, Benjamin T.; Matson, Cole W.; Aiken, George R.; Hsu-Kim, Heileen

2017-01-01

The toxicity of soluble metal-based nanomaterials may be due to the uptake of metals in both dissolved and nanoparticulate forms, but the relative contributions of these different forms to overall metal uptake rates under environmental conditions are not quantitatively defined. Here, we investigated the linkage between the dissolution rates of copper(II) oxide (CuO) nanoparticles (NPs) and their bioavailability to Gulf killifish (Fundulus grandis) embryos, with the aim of quantitatively delineating the relative contributions of nanoparticulate and dissolved species for Cu uptake. Gulf killifish embryos were exposed to dissolved Cu and CuO NP mixtures comprising a range of pH values (6.3–7.5) and three types of natural organic matter (NOM) isolates at various concentrations (0.1–10 mg-C L–1), resulting in a wide range of CuO NP dissolution rates that subsequently influenced Cu uptake. First-order dissolution rate constants of CuO NPs increased with increasing NOM concentration and for NOM isolates with higher aromaticity, as indicated by specific ultraviolet absorbance (SUVA), while Cu uptake rate constants of both dissolved Cu and CuO NP decreased with NOM concentration and aromaticity. As a result, the relative contribution of dissolved Cu and nanoparticulate CuO species for the overall Cu uptake rate was insensitive to NOM type or concentration but largely determined by the percentage of CuO that dissolved. These findings highlight SUVA and aromaticity as key NOM properties affecting the dissolution kinetics and bioavailability of soluble metal-based nanomaterials in organic-rich waters. These properties could be used in the incorporation of dissolution kinetics into predictive models for environmental risks of nanomaterials.

NORMAL NASAL GENE EXPRESSION LEVELS USING CDNA ARRAY TECHNOLOGY

Science.gov (United States)

Normal Nasal Gene Expression Levels Using cDNA Array Technology. The nasal epithelium is a target site for chemically-induced toxicity and carcinogenicity. To detect and analyze genetic events which contribute to nasal tumor development, we first defined the gene expressi...

Annual increments, specific gravity and energy of Eucalyptus grandis by gamma-ray attenuation technique

International Nuclear Information System (INIS)

Rezende, M.A.; Guerrini, I.A.; Ferraz, E.S.B.

1990-01-01

Specific gravity annual increments in volume, mass and energy of Eucalyptus grandis at thirteen years of age were made taking into account measurements of the calorific value for wood. It was observed that the calorific value for wood decrease slightly, while the specific gravity increase significantly with age. The so-called culmination age for the Annual Volume Increment was determined to be around fourth year of growth while for the Annual Mass and Energy Increment was around the eighty year. These results show that a tree in a particular age may not have a significant growth in volume, yet one is mass and energy. (author)

Molecular cloning and sequence analysis of hamster CENP-A cDNA

Directory of Open Access Journals (Sweden)

Valdivia Manuel M

2002-05-01

Full Text Available Abstract Background The centromere is a specialized locus that mediates chromosome movement during mitosis and meiosis. This chromosomal domain comprises a uniquely packaged form of heterochromatin that acts as a nucleus for the assembly of the kinetochore a trilaminar proteinaceous structure on the surface of each chromatid at the primary constriction. Kinetochores mediate interactions with the spindle fibers of the mitotic apparatus. Centromere protein A (CENP-A is a histone H3-like protein specifically located to the inner plate of kinetochore at active centromeres. CENP-A works as a component of specialized nucleosomes at centromeres bound to arrays of repeat satellite DNA. Results We have cloned the hamster homologue of human and mouse CENP-A. The cDNA isolated was found to contain an open reading frame encoding a polypeptide consisting of 129 amino acid residues with a C-terminal histone fold domain highly homologous to those of CENP-A and H3 sequences previously released. However, significant sequence divergence was found at the N-terminal region of hamster CENP-A that is five and eleven residues shorter than those of mouse and human respectively. Further, a human serine 7 residue, a target site for Aurora B kinase phosphorylation involved in the mechanism of cytokinesis, was not found in the hamster protein. A human autoepitope at the N-terminal region of CENP-A described in autoinmune diseases is not conserved in the hamster protein. Conclusions We have cloned the hamster cDNA for the centromeric protein CENP-A. Significant differences on protein sequence were found at the N-terminal tail of hamster CENP-A in comparison with that of human and mouse. Our results show a high degree of evolutionary divergence of kinetochore CENP-A proteins in mammals. This is related to the high diverse nucleotide repeat sequences found at the centromere DNA among species and support a current centromere model for kinetochore function and structural

ESTs, cDNA microarrays, and gene expression profiling: tools for dissecting plant physiology and development.

Science.gov (United States)

Alba, Rob; Fei, Zhangjun; Payton, Paxton; Liu, Yang; Moore, Shanna L; Debbie, Paul; Cohn, Jonathan; D'Ascenzo, Mark; Gordon, Jeffrey S; Rose, Jocelyn K C; Martin, Gregory; Tanksley, Steven D; Bouzayen, Mondher; Jahn, Molly M; Giovannoni, Jim

2004-09-01

Gene expression profiling holds tremendous promise for dissecting the regulatory mechanisms and transcriptional networks that underlie biological processes. Here we provide details of approaches used by others and ourselves for gene expression profiling in plants with emphasis on cDNA microarrays and discussion of both experimental design and downstream analysis. We focus on methods and techniques emphasizing fabrication of cDNA microarrays, fluorescent labeling, cDNA hybridization, experimental design, and data processing. We include specific examples that demonstrate how this technology can be used to further our understanding of plant physiology and development (specifically fruit development and ripening) and for comparative genomics by comparing transcriptome activity in tomato and pepper fruit.

Generation of a reliable full-length cDNA of infectiousTembusu virus using a PCR-based protocol.

Science.gov (United States)

Liang, Te; Liu, Xiaoxiao; Cui, Shulin; Qu, Shenghua; Wang, Dan; Liu, Ning; Wang, Fumin; Ning, Kang; Zhang, Bing; Zhang, Dabing

2016-02-02

Full-length cDNA of Tembusu virus (TMUV) cloned in a plasmid has been found instable in bacterial hosts. Using a PCR-based protocol, we generated a stable full-length cDNA of TMUV. Different cDNA fragments of TMUV were amplified by reverse transcription (RT)-PCR, and cloned into plasmids. Fragmented cDNAs were amplified and assembled by fusion PCR to produce a full-length cDNA using the recombinant plasmids as templates. Subsequently, a full-length RNA was transcribed from the full-length cDNA in vitro and transfected into BHK-21 cells; infectious viral particles were rescued successfully. Following several passages in BKH-21 cells, the rescued virus was compared with the parental virus by genetic marker checks, growth curve determinations and animal experiments. These assays clearly demonstrated the genetic and biological stabilities of the rescued virus. The present work will be useful for future investigations on the molecular mechanisms involved in replication and pathogenesis of TMUV. Copyright © 2015 Elsevier B.V. All rights reserved.

EFFECT OF THE STEAMING ON THE DRYING OF Eucalyptus grandis BOARDS

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Raphael Nogueira Rezende

2015-03-01

Full Text Available The objective of this work was to evaluate the effect of the steaming on the drying rate and drying quality of Eucalyptus grandis boards. For this purpose, wood from an experimental plantation of the Federal University of Lavras, Minas Gerais State, Brazil, with 24 years of age was used. Trees were felled and sectioned in logs and the logs were sawn by a tangential system. Half of the boards volume were steamed during 3 hours at 90ºC of temperature and 100% of relative humidity after the heating of the drying process. The other half was not steamed (control. The boards were dried in the dry-kiln and the resulting defects from the drying process and drying rate were determined. The results indicated that the steaming was effective in increase of the drying rate in 15% and decrease of the drying defects of 20 to 52%.

Expression of wild-type and mutant medium-chain acyl-CoA dehydrogenase (MCAD) cDNA in eucaryotic cells

DEFF Research Database (Denmark)

Jensen, T G; Andresen, B S; Bross, P

1992-01-01

An effective EBV-based expression system for eucaryotic cells has been developed and used for the study of the mitochondrial enzyme medium-chain acyl-CoA dehydrogenase (MCAD). 1325 bp of PCR-generated MCAD cDNA, containing the entire coding region, was placed between the SV40 early promoter...... and polyadenylation signals in the EBV-based vector. Both wild-type MCAD cDNA and cDNA containing the prevalent disease-causing mutation A to G at position 985 of the MCAD cDNA were tested. In transfected COS-7 cells, the steady state amount of mutant MCAD protein was consistently lower than the amount of wild......-type human enzyme. The enzyme activity in extracts from cells harbouring the wild-type MCAD cDNA was dramatically higher than in the controls (harbouring the vector without the MCAD gene) while only a slightly higher activity was measured with the mutant MCAD. The mutant MCAD present behaves like wild...

ESPECTRORRADIOMETRIA NA REGIÃO DO VISÍVEL E DO INFRAVERMELHO PRÓXIMO EM POVOAMENTO DE Eucalyptus grandis Hill ex Maiden

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Catize Brandelero

2012-01-01

Full Text Available Reflectance readings in border and inner tree leaves in a Eucalyptus grandis stand, in São Pedro das Missões, Rio Grande do Sul state, were analyzed in the regions of the visible electromagnetic spectrum and the nearby infrared, by using spectrum radiometry. The area was divided in two parts: border and center stands. In order to collect the material, the crown was divided in three parts (superior, medium and inferior, so that it would be possible to differentiate the positions of leaf collections in each area. Three trees were sampled in each area, adding up to six trees, for each tree, 60 isolated leaves were collected, 20 in each position. The reflectance readings were carried out through FieldSpec®3 spectrum radiometer and the final results were segmented in the visible and nearby infrared spectral bands. The statistical analysis was made on the basis of several tests, among them Tukey HSD test, in order to compare the averages of the visible region, which, according to ANOVA, present significant differences. It is concluded that the collecting indicating class of leaves for the spectrum radiometric analysis in the visible region are preferably the 5 one (tree in the center, reading in the medium part and #3 one (border tree, reading superior part.

Isolation and Expression analysis of OsPME1, encoding for a putative Pectin Methyl Esterase from Oryza sativa (subsp. indica)

OpenAIRE

Kanneganti, Vydehi; Gupta, Aditya Kumar

2009-01-01

Pectin Methyl Esterases (PMEs) play an essential role during plant development by affecting the mechanical properties of the plant cell walls. Recent studies indicated that PMEs play important role in pollen tube development. In this study, we isolated a 1.3 kb cDNA clone from rice panicle cDNA library. It contained a 1038 bp of open reading frame (ORF) encoding for a putative pectin methyl esterase of 345 aminoacids with a 20 aminoacid signal peptide and was hence designated as OsPME1 (Oryza...

RFLP for Duchenne muscular dystrophy cDNA clone 30-2

Energy Technology Data Exchange (ETDEWEB)

Walker, A P; Bartlett, R J; Laing, N G; Siddique, T; Yamaoka, L H; Chen, J C; Hung, W Y; Roses, A D [Duke Univ. Medical Center, Durham, NC (USA)

1988-09-26

30-2 is one of 6 cDNA clones which comprise the cDNA for the Duchenne muscular dystrophy gene. It is a 1.15 kb fragment in the EcoRI site of Bluescribe. TaqI (T{down arrow}CGA) identifies two bands with alleles at 3.7 and 3.5 kb, as well as eight constant bands at 9.0, 7.5, 4.6, 3.6, 3.4, 2.5, 1.7 and 1.4 kb. The allele frequency was studied in 47 unrelated DMD males: 3.7 kb allele 0.45; and 3.5 kb allele 0.55. Co-dominant X-linked segregation was demonstrated in two 2-generation families. 1.1% agarose gels required to resolve the bands. The polymorphism is also recognized by PERT 87-15.

Organogênese de explante foliar de clones de Eucalyptus grandis x E. urophylla Organogenesis of the leaf explant of Eucalyptus grandis x E. urophylla clones

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Elisa Cristina Soares de Carvalho Alves

2004-05-01

Full Text Available O objetivo deste trabalho foi avaliar os efeitos dos reguladores de crescimento TDZ [1-fenil-3-(1,2,3-tia-diazol-5-iluréia], BAP (6-benzilaminopurina e ANA (ácido naftalenoacético no desempenho da propagação in vitro por organogênese de explante foliar de três clones híbridos de Eucalyptus grandis x Eucalyptus urophylla. Houve resposta diferenciada dos clones quanto a intensidade, textura e coloração dos calos, em razão dos tratamentos com os reguladores de crescimento. Os melhores resultados de calejamento dos três genótipos foram observados nos tratamentos com a combinação dos reguladores de crescimento TDZ (0,5 mg L-1 e ANA (0,1 mg L-1, obtendo-se 100% de calejamento no explante foliar. Os piores resultados de calejamento foram observados nos tratamentos com a combinação dos reguladores de crescimento BAP (0,1 mg L-1 e ANA (0,1 mg L-1. Em relação à regeneração, a melhor resposta foi obtida com 1,0 mg L-1 BAP em que 8% dos calos formados a partir de explantes foliares regeneraram gemas, com número médio destas formadas por calo igual a 4,2.The aim of this work was to evaluate the effects of growth regulators TDZ [1-phenil-3-(1,2,3-thiadiazol-5-yl urea], BAP (6-benzilaminopurine e NAA (Naphthalene acetic acid on the in vitro propagation by organogenesis from foliar explants of Eucalyptus grandis x E. urophylla. Depending on the clone used, there were singular responses to growth regulators treatment regarding callusing intensity, texture and color. The best results of the three genotypes used were observed with the TDZ (0.5 mg L-1 and NAA (0.1 mg L-1 treatment, where 100% of the foliar explants presented callus. The worst results were observed with the BAP (0.1 mg L-1 and NAA (0.1 mg L-1 treatment. Subsequently, considering the regeneration process, the best response was achieved with 1.0 mg L-1 BAP, in which 8% of the calli regenerated buds, with an average of 4.2 buds per explant.

Effect of Different Lignocellulosic Diets on Bacterial Microbiota and Hydrolytic Enzyme Activities in the Gut of the Cotton Boll Weevil (Anthonomus grandis).

Science.gov (United States)

Ben Guerrero, Emiliano; Soria, Marcelo; Salvador, Ricardo; Ceja-Navarro, Javier A; Campos, Eleonora; Brodie, Eoin L; Talia, Paola

2016-01-01

Cotton boll weevils, Anthonomus grandis , are omnivorous coleopteran that can feed on diets with different compositions, including recalcitrant lignocellulosic materials. We characterized the changes in the prokaryotic community structure and the hydrolytic activities of A. grandis larvae fed on different lignocellulosic diets. A. grandis larvae were fed on three different artificial diets: cottonseed meal (CM), Napier grass (NG) and corn stover (CS). Total DNA was extracted from the gut samples for amplification and sequencing of the V3-V4 hypervariable region of the 16S rRNA gene. Proteobacteria and Firmicutes dominated the gut microbiota followed by Actinobacteria, Spirochaetes and a small number of unclassified phyla in CM and NG microbiomes. In the CS feeding group, members of Spirochaetes were the most prevalent, followed by Proteobacteria and Firmicutes. Bray-Curtis distances showed that the samples from the CS community were clearly separated from those samples of the CM and NG diets. Gut extracts from all three diets exhibited endoglucanase, xylanase, β-glucosidase and pectinase activities. These activities were significantly affected by pH and temperature across different diets. We observed that the larvae reared on a CM showed significantly higher activities than larvae reared on NG and CS. We demonstrated that the intestinal bacterial community structure varies depending on diet composition. Diets with more variable and complex compositions, such as CS, showed higher bacterial diversity and richness than the two other diets. In spite of the detected changes in composition and diversity, we identified a core microbiome shared between the three different lignocellulosic diets. These results suggest that feeding with diets of different lignocellulosic composition could be a viable strategy to discover variants of hemicellulose and cellulose breakdown systems.

Evaluación de la toxicidad de proteínas de Bacillus thuringiensis Berliner sobre el picudo del algodonero Anthonomus grandis Boheman

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Gómez Sylvia

2006-12-01

Full Text Available

El picudo del algodonero, Anthonomus grandis Boheman (Coleoptera: Curculionidae, es considerado la principal plaga del cultivo de algodón en Colombia, no sólo por los daños económicos y sociales que causa, sino porque con sus apariciones tempranas altera e interrumpe el desarrollo de los programas de manejo integrado de plagas. La importancia de un manejo inteligente radica en el hecho de que este insecto se  comporta como plaga ‘clave’, con una capacidad de daño en el cultivo entre 50% y 90%, si no se controla. En el presente estudio se estableció una metodología de bioensayo que determinó la actividad tóxica de  proteínas Cry3Aa y Cry1Ia de cepas de B. thuringiensis sobre larvas de primer instar de A. grandis. En los bioensayos se empleó una dieta artificial mezclada con los extractos bacterianos que contenían el complejo espora-cristal de cada cepa. Los resultados indicaron que la cepa de referencia variedad san diego presentó toxicidad sobre larvas de primer instar de A. grandis, en condiciones de laboratorio, con una concentración letal 50 (CL50 de 147,6143 μg de proteína total por mililitro de volumen final de dieta artificial, representando una alternativa potencial para el manejo de este insecto plaga del cultivo de algodón.

Good quality Vitis RNA obtained from an adapted DNA isolation protocol

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Isabel Baiges

2003-03-01

Full Text Available Grapevine is a woody plant, whose high carbohydrate and phenolic compound contents usually interferes with nucleic acid isolation. After we tried several protocols for isolating RNA from the Vitis rootstock Richter- 110 (R-110 with little or no success, we adapted a method reported to be satisfactory for grapevine DNA isolation, to extract RNA. With slight protocol modifications, we succeeded to obtain polysaccharide- and phenolic-free RNA preparations from all vegetative tissues, without excessive sample handling. RNA isolated by the reported method permitted to obtain highly pure mRNA (messenger RNA to construct a cDNA (complementary DNA library and allowed gene transcription analysis by reverse Northern, which guarantees RNA integrity. This method may also be suitable for other plant species with high polysaccharide or phenolic contents.

CARACTERÍSTICAS TECNOLÓGICAS DAS MADEIRAS DE Eucalyptus grandis W.Hill ex Maiden E Eucalyptus cloeziana F. Muell VISANDO AO SEU APROVEITAMENTO NA INDÚSTRIA MOVELEIRA

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Ailton Teixeira do Vale

2006-07-01

Full Text Available Este trabalho se desenvolveu na Universidade de Brasília e no Laboratório de Produtos Florestais (IBAMA, Brasília, DF. Foram estudadas duas espécies de eucalipto (Eucalyptus grandis W.Hill ex Maiden e Eucalyptus cloeziana para confecção de peças mobiliárias. A madeira de E. grandis apresenta propriedades físicas (densidade e retratibilidade e mecânicas (flexão estática e dureza extremamente positivas para a indústria moveleira, sendo complementado por seu bom desempenho perante equipamentos e máquinas, além de receber bem produtos de acabamento. A cor da madeira e o seu desenho levaram os consumidores a mostrar ótima aceitação do móvel fabricado com a espécie. A madeira de Eucalyptus cloeziana, apesar de mostrar propriedades físicas e mecânicas com valores mais elevados que as do Eucalyptus grandis W.Hill ex Maiden, apresenta características desejadas para indústria moveleira. A sua coloração cinza oliva é uma opção para o consumidor. Alguns cuidados especiais com essa espécie deverão ser tomados durante operações com máquinas e equipamentos. Os valores da propriedade dureza apresentados por essa madeira a indicam para fabricação de piso.

Isolation of high-quality total RNA from leaves of Myrciaria dubia "CAMU CAMU".

Science.gov (United States)

Gómez, Juan Carlos Castro; Reátegui, Alina Del Carmen Egoavil; Flores, Julián Torres; Saavedra, Roberson Ramírez; Ruiz, Marianela Cobos; Correa, Sixto Alfredo Imán

2013-01-01

Myrciaria dubia is a main source of vitamin C for people in the Amazon region. Molecular studies of M. dubia require high-quality total RNA from different tissues. So far, no protocols have been reported for total RNA isolation from leaves of this species. The objective of this research was to develop protocols for extracting high-quality total RNA from leaves of M. dubia. Total RNA was purified following two modified protocols developed for leaves of other species (by Zeng and Yang, and by Reid et al.) and one modified protocol developed for fruits of the studied species (by Silva). Quantity and quality of purified total RNA were assessed by spectrophotometric and electrophoretic analysis. Additionally, quality of total RNA was evaluated with reverse-transcription polymerase chain reaction (RT-PCR). With these three modified protocols we were able to isolate high-quality RNA (A260nm/A280nm >1.9 and A260nm/A230nm >2.0). Highest yield was produced with the Zeng and Yang modified protocol (384±46µg ARN/g fresh weight). Furthermore, electrophoretic analysis showed the integrity of isolated RNA and the absence of DNA. Another proof of the high quality of our purified RNA was the successful cDNA synthesis and amplification of a segment of the M. dubia actin 1 gene. We report three modified protocols for isolation total RNA from leaves of M. dubia. The modified protocols are easy, rapid, low in cost, and effective for high-quality and quantity total RNA isolation suitable for cDNA synthesis and polymerase chain reaction.

Multiplex cDNA quantification method that facilitates the standardization of gene expression data

Science.gov (United States)

Gotoh, Osamu; Murakami, Yasufumi; Suyama, Akira

2011-01-01

Microarray-based gene expression measurement is one of the major methods for transcriptome analysis. However, current microarray data are substantially affected by microarray platforms and RNA references because of the microarray method can provide merely the relative amounts of gene expression levels. Therefore, valid comparisons of the microarray data require standardized platforms, internal and/or external controls and complicated normalizations. These requirements impose limitations on the extensive comparison of gene expression data. Here, we report an effective approach to removing the unfavorable limitations by measuring the absolute amounts of gene expression levels on common DNA microarrays. We have developed a multiplex cDNA quantification method called GEP-DEAN (Gene expression profiling by DCN-encoding-based analysis). The method was validated by using chemically synthesized DNA strands of known quantities and cDNA samples prepared from mouse liver, demonstrating that the absolute amounts of cDNA strands were successfully measured with a sensitivity of 18 zmol in a highly multiplexed manner in 7 h. PMID:21415008

cDNA cloning, sequence analysis, and chromosomal localization of the gene for human carnitine palmitoyltransferase

International Nuclear Information System (INIS)

Finocchiaro, G.; Taroni, F.; Martin, A.L.; Colombo, I.; Tarelli, G.T.; DiDonato, S.; Rocchi, M.

1991-01-01

The authors have cloned and sequenced a cDNA encoding human liver carnitine palmitoyltransferase an inner mitochondrial membrane enzyme that plays a major role in the fatty acid oxidation pathway. Mixed oligonucleotide primers whose sequences were deduced from one tryptic peptide obtained from purified CPTase were used in a polymerase chain reaction, allowing the amplification of a 0.12-kilobase fragment of human genomic DNA encoding such a peptide. A 60-base-pair (bp) oligonucleotide synthesized on the basis of the sequence from this fragment was used for the screening of a cDNA library from human liver and hybridized to a cDNA insert of 2255 bp. This cDNA contains an open reading frame of 1974 bp that encodes a protein of 658 amino acid residues including 25 residues of an NH 2 -terminal leader peptide. The assignment of this open reading frame to human liver CPTase is confirmed by matches to seven different amino acid sequences of tryptic peptides derived from pure human CPTase and by the 82.2% homology with the amino acid sequence of rat CPTase. The NH 2 -terminal region of CPTase contains a leucine-proline motif that is shared by carnitine acetyl- and octanoyltransferases and by choline acetyltransferase. The gene encoding CPTase was assigned to human chromosome 1, region 1q12-1pter, by hybridization of CPTase cDNA with a DNA panel of 19 human-hanster somatic cell hybrids

Filtration and clearance rates of Anadara grandis juveniles (Pelecypoda, Arcidae with different temperatures and suspended matter concentrations

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Anselmo Miranda-Baeza

2006-09-01

Full Text Available The mangrove cockle Anadara grandis (Broderip and Sowerby, 1829 is a potential candidate for aquaculture and for bioremediation of aquaculture effluents in the tropical and subtropical coastal areas of the eastern Pacific Ocean. Laboratory-produced spat are available, but there is no information on their responses to the range of environmental conditions to which they might be subject during the growth cycle. The aim of this study was to evaluate the filtration and clearance rates of A. grandis spat (shell length 9.50±0.37 mm with a food concentration (7.5 mg∙l-1 at four different temperatures (22, 25, 28 and 31 ºC, with pH = 7.5±0.2 and O2 concentration of 6.4±0.5 mg·l-1; experiment one; and with a temperature (25 °C and five concentrations of suspended matter (from 7.5 to 29 mg·l-1 and pH and O2 values of 7.9±0.2 and 6.8± 0.4 mg·l-1; experiment two. Filtration and clearance rates were highest at 25 ºC and significantly different (p.05. In the second experiment filtration increased according to the amount of food available, but there were no significant differences (p>.05 between 7.5 and 11 mg·l-1 and from 22.4 to 29 mg·l-1. The trend was similar for clearance, and in this case significant differences were found (pLa almeja Anadara grandis (Broderip and Sowerby, 1829 es un candidato potencial para la acuicultura y la biorremediación de efluentes acuícolas en las áreas costeras tropicales y subtropicales del océano Pacífico oriental. Se dispone de semilla producida en laboratorio, sin embargo no hay información sobre sus respuestas a los intervalos de las condiciones ambientales a las cuales puede estar sujeta durante el periodo de crecimiento. El objetivo de este estudio fue evaluar las tasas de filtración y de clarificación de semilla de A. grandis (largo de la concha= 9.50±0.37 mm con una concentración de alimento (7.5 mg∙l-1 y cuatro diferentes temperaturas (22, 25, 28 y 31 °C con pH= 7.5±0.2, concentración de O

Cloning of soluble alkaline phosphatase cDNA and molecular basis of the polymorphic nature in alkaline phosphatase isozymes of Bombyx mori midgut.

Science.gov (United States)

Itoh, M; Kanamori, Y; Takao, M; Eguchi, M

1999-02-01

A cDNA coding for soluble type alkaline phosphatase (sALP) of Bombyx mori was isolated. Deduced amino acid sequence showed high identities to various ALPs and partial similarities to ATPase of Manduca sexta. Using this cDNA sequence as a probe, the molecular basis of electrophoretic polymorphism in sALP and membrane-bound type ALP (mALP) was studied. As for mALP, the result suggested that post-translational modification was important for the proteins to express activity and to represent their extensive polymorphic nature, whereas the magnitude of activities was mainly regulated by transcription. On the other hand, sALP zymogram showed poor polymorphism, but one exception was the null mutant, in which the sALP gene was largely lost. Interestingly, the sALP gene was shown to be transcribed into two mRNAs of different sizes, 2.0 and 2.4 Kb. In addition to the null mutant of sALP, we found a null mutant for mALP. Both of these mutants seem phenotypically silent, suggesting that the functional differentiation between these isozymes is not perfect, so that they can still work mutually and complement each other as an indispensable enzyme for B. mori.

Selection of Bacillus thuringiensis strains toxic to cotton boll weevil (Anthonomus grandis, Coleoptera: Curculionidae) larvae.

Science.gov (United States)

Pérez, Melisa P; Sauka, Diego H; Onco, María I; Berretta, Marcelo F; Benintende, Graciela B

Preliminary bioassays with whole cultures (WC) of 124 Bacillus thuringiensis strains were performed with neonate larvae of Anthonomus grandis, a major cotton pest in Argentina and other regions of the Americas. Three exotic and four native strains were selected for causing more than 50% mortality. All of them were β-exotoxin producers. The native strains shared similar morphology of parasporal crystals, similar protein pattern and identical insecticidal gene profiles. These features resembled Lepidoptera-toxic strains. Furthermore, these strains showed a Rep-PCR pattern identical to lepidoptericidal strain HD-1, suggesting that these strains may belong to serovar kurstaki. However, some differences were observed in the plasmid profiles and in the production of β-exotoxin. To determine the culture fractions where the insecticidal metabolites were present, bioassays including resuspended spore-crystal pellets, filtered supernatants (FS) were compared with those of WC. Both fractions tested showed some level of insecticidal activity. The results may suggest that the main toxic factors can be found in FS and could be directly correlated with the presence of β-exotoxin. Based on the bioassays with FS and autoclaved FS, the participation of thermolabile virulence factors such as Cry1I in toxicity is neither discarded. In the selected strains, β-exotoxin would be the major associated virulence factor; therefore, their use in biological control of A. grandis should be restricted. Nevertheless, these strains could be the source of genes (e.g., cry1Ia) to produce transgenic cotton plants resistant to this pest. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Isolation and expression pattern of COR15b and KIN1 genes in ...

African Journals Online (AJOL)

STORAGESEVER

2009-11-02

Nov 2, 2009 ... COR15b and KIN1 (COR 6.5) genes encode polypeptides of 15 KDa and 6.5 KDa, respectively. They are involved in the dehydration tolerance mechanisms and play important role under cold stress. cDNA sequences of COR15b and KIN1 genes were firstly isolated from leaves of watermelon (Citrullus.

Full-length cDNA sequences from Rhesus monkey placenta tissue: analysis and utility for comparative mapping

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Lee Sang-Rae

2010-07-01

Full Text Available Abstract Background Rhesus monkeys (Macaca mulatta are widely-used as experimental animals in biomedical research and are closely related to other laboratory macaques, such as cynomolgus monkeys (Macaca fascicularis, and to humans, sharing a last common ancestor from about 25 million years ago. Although rhesus monkeys have been studied extensively under field and laboratory conditions, research has been limited by the lack of genetic resources. The present study generated placenta full-length cDNA libraries, characterized the resulting expressed sequence tags, and described their utility for comparative mapping with human RefSeq mRNA transcripts. Results From rhesus monkey placenta full-length cDNA libraries, 2000 full-length cDNA sequences were determined and 1835 rhesus placenta cDNA sequences longer than 100 bp were collected. These sequences were annotated based on homology to human genes. Homology search against human RefSeq mRNAs revealed that our collection included the sequences of 1462 putative rhesus monkey genes. Moreover, we identified 207 genes containing exon alterations in the coding region and the untranslated region of rhesus monkey transcripts, despite the highly conserved structure of the coding regions. Approximately 10% (187 of all full-length cDNA sequences did not represent any public human RefSeq mRNAs. Intriguingly, two rhesus monkey specific exons derived from the transposable elements of AluYRa2 (SINE family and MER11B (LTR family were also identified. Conclusion The 1835 rhesus monkey placenta full-length cDNA sequences described here could expand genomic resources and information of rhesus monkeys. This increased genomic information will greatly contribute to the development of evolutionary biology and biomedical research.

EFFECT OF ARBUSCULAR MYCORRHIZA FUNGI INOCULATION ON TEAK (Tectona grandis Linn. F AT CIKAMPEK, WEST JAVA

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Ragil S.B. Irianto

2005-07-01

Full Text Available The aim of this study was to identify the effect of Arbuscular Mycorhiza Fungi (AMF on the early growth of teak (Tectona grandis Linn. F plantation. Teak seedlings were inoculated with Glomus aggregatum or Mycofer (mixing of four Arbuscular Mycorrhiza Fungi (AMF : G. margarita, G. manihotis, G. etunicatum and Acalospora spinosa at the time of transplantation. At  three months old the seedlings were planted in Cikampek experimental forest. Results showed that application of G. aggregatum or mycofer to teak could accelerate height and diameter growth by up to 61%and4 7%, respectively, after three months in the field.

Leaf area index estimation of Eucalyptus grandis W.Hill. in plantations

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Dubal Papamija-Muñoz

2012-12-01

Full Text Available We estimated leaf area index (LAI in Eucalyptus grandis W.Hill. plantations in four farms in the Smurfit Kappa Carton de Colombia (SKCC with three farms located in the city of Popayan (Cauca and one located in the municipality of Restrepo (Valle del Cauca. Each farm had three fertilized and three unfertilized plots with 64 individuals in each. We used three methods, Plant Canopy Analyzer 2000 (PCA 2000, flat photograph PIPEcv software and a destructive method, which was generated using a mathematical model. The first two methods were measured bimonthly for a year and the final method required trees being cut to measure their diameter. Estimation of leaf area index was 2.01 for PCA 2000, 3.12 for PIPEcv and 2.83 for the mathematical model. These values correspond to the average and range of leaf area indices obtained for each method on all farms. Statistically the three methodologies developed in this study were not closely related.

Characterization of the cDNA encoding human nucleophosmin and studies of its role in normal and abnormal growth

International Nuclear Information System (INIS)

Chan, Waiyee; Liu, Qingrong; Borjigin, J.; Busch, H.; Rennert, O.M.; Tease, L.A.; Chan, Puikwong

1989-01-01

A cDNA encoding human nucleophosmin (protein B23) was obtained by screening a human placental cDNA library in δgtll first with monoclonal antibody to rat nucleophosmin and then with confirmed partial cDNA of human nucleophosmin as probes. The cDNA had 1,311 bp with a coding sequence encoding a protein of 294 amino acids. The identity of the cDNA was confirmed by the presence of encoded amino acid sequences identical with those determined by sequencing pure rat nucleophosmin (a total of 138 amino acids). The most striking feature of the sequence is an acidic cluster located in the middle of the molecule. The cluster consists of 26 Asp/Glu and 1 Phe and Ala. Comparison of human nucleophosmin and Xenopus nucleolar protein NO38 shows 64.3% sequence identity. The N-terminal 130 amino acids of human nucleophosmin also bear 50% identity with that of Xenopus nucleoplasmin. Northern blot analysis of rat liver total RNA with a partial nucleophosmin cDNA as probe demonstrated a homogeneous mRNA band of about 1.6 kb. Similar observations were made in hypertrophic rat liver and Novikoff hepatoma. When the protein levels were compared with Western blot immunoassays, Navikoff hepatoma showed 20 times more nucleophosmin, while only about 5 times more nucleophosmin was observed in hypertrophic rat liver than in unstimulated normal liver

Characterization and immunological identification of cDNA clones encoding two human DNA topoisomerase II isozymes

International Nuclear Information System (INIS)

Chung, T.D.Y.; Drake, F.H.; Tan, K.B.; Per, S.R.; Crooke, S.T.; Mirabelli, C.K.

1989-01-01

Several DNA topoisomerase II partial cDNA clones obtained from a human Raji-HN2 cDNA library were sequenced and two classes of nucleotide sequences were found. One member of the first class, SP1, was identical to an internal fragment of human HeLa cell Topo II cDNA described earlier. A member of the second class, SP11, shared extensive nucleotide (75%) and predicted peptide (92%) sequence similarities with the first two-thirds of HeLa Topo II. Each class of cDNAs hybridized to unique, nonoverlapping restriction enzyme fragments of genomic DNA from several human cell lines. Synthetic 24-mer oligonucleotide probes specific for each cDNA class hybridized to 6.5-kilobase mRNAs; furthermore, hybridization of probe specific for one class was not blocked by probe specific for the other. Antibodies raised against a synthetic SP1-encoded dodecapeptide specifically recognized the 170-kDa form of Topo II, while antibodies raised against the corresponding SP11-encoded dodecapeptide, or a second unique SP11-encoded tridecapeptide, selectively recognized the 180-kDa form of Topo II. These data provide genetic and immunochemical evidence for two Topo II isozymes

Purification, cDNA cloning and modification of a defensin from the coconut rhinoceros beetle, Oryctes rhinoceros.

Science.gov (United States)

Ishibashi, J; Saido-Sakanaka, H; Yang, J; Sagisaka, A; Yamakawa, M

1999-12-01

A novel member of the insect defensins, a family of antibacterial peptides, was purified from larvae of the coconut rhinoceros beetle, Oryctes rhinoceros, immunized with Escherichia coli. A full-size cDNA was cloned by combining reverse-transcription PCR (RT-PCR), and 5'- and 3'-rapid amplification of cDNA ends (RACE). Analysis of the O. rhinoceros defensin gene expression showed it to be expressed in the fat body and hemocyte, midgut and Malpighian tubules. O. rhinoceros defensin showed strong antibacterial activity against Staphylococcus aureus. A 9-mer peptide amidated at its C-terminus, AHCLAICRK-NH2 (Ala22-Lys30-NH2), was synthesized based on the deduced amino-acid sequence, assumed to be an active site sequence by analogy with the sequence of a defensin isolated from larvae of the beetle Allomyrina dichotoma. This peptide showed antibacterial activity against S. aureus, methicillin-resistant S. aureus, E. coli and Pseudomonas aeruginosa. We further modified this oligopeptide and synthesized five 9-mer peptides, ALRLAIRKR-NH2, ALLLAIRKR-NH2, AWLLAIRKR-NH2, ALYLAIRKR-NH2 and ALWLAIRKR-NH2. These oligopeptides showed strong antibacterial activity against Gram-negative and Gram-positive bacteria. The antibacterial effect of Ala22-Lys30-NH2 analogues was due to its interaction with bacterial membranes, judging from the leakage of liposome-entrapped glucose. These Ala22-Lys30-NH2 analogues did not show haemolytic activity and did not inhibit the growth of murine fibroblast cells or macrophages, except for AWLLAIRKR-NH2.

cDNA, genomic cloning and sequence analysis of ribosomal protein ...

African Journals Online (AJOL)

enoh

2012-03-13

Mar 13, 2012 ... cDNA and the genomic sequence of RPS4X were cloned successfully from ... S4 genes plays a role in Turner syndrome; however, this ..... Project of Educational Committee of Sichuan Province ... Molecular biology of the cell.

First occurrence of Alcaeorrhynchus grandis (Dallas) (Hemiptera: Pentatomidae) preying on defoliating caterpillars of oil palm in the state of Para, Brazil; Primeira ocorrencia de Alcaeorrhynchus grandis (Dallas) (Hemiptera: Pentatomidae) predando lagartas desfolhadoras do dendezeiro no estado do Para, Brasil

Energy Technology Data Exchange (ETDEWEB)

Ribeiro, Rafael C.; Lemos, Walkymario P.; Muller, Antonio A. [EMBRAPA Amazonia Oriental, Belem, PA (Brazil). Lab. de Entomologia], e-mail: rafaufra@yahoo.com.br, e-mail: wplemos@cpatu.embrapa.br; Muller, Antonio A. [Embrapa Amazonia Oriental, Belem, PA (Brazil). Lab. de Entomologia; Bernardino, Aline S.; Buecke, Joel [Grupo Agropalma S/A., Tailandia, PA (Brazil)

2010-01-15

The oil palm Elaeis guineensis is usually attacked by pests, particularly, defoliating caterpillars. Between 2004 and 2006 a stinkbug predator (Asopinae) was registered preying on caterpillars of Brassolis sophorae L., Opsiphanes invirae Hubner (Lepidoptera: Nymphalidae) and Sibine spp. (Lepidoptera: Limacodidae), reducing their populations in commercial oil palm plantations in the State of Para, Brazil. Specimens of the natural enemy were collected, mounted, and identified as Alcaeorrhynchus grandis (Dallas) (Hemiptera: Pentatomidae), corresponding to the first report of the occurrence of this stinkbug attacking defoliating caterpillars of oil palm in Brazil. (author)

Genome-Wide Analysis of the AP2/ERF Family in Eucalyptus grandis: An Intriguing Over-Representation of Stress-Responsive DREB1/CBF Genes

Science.gov (United States)

SanClemente, H.; Mounet, F.; Dunand, C.; Marque, G.; Marque, C.; Teulières, C.

2015-01-01

Background The AP2/ERF family includes a large number of developmentally and physiologically important transcription factors sharing an AP2 DNA-binding domain. Among them DREB1/CBF and DREB2 factors are known as master regulators respectively of cold and heat/osmotic stress responses. Experimental Approaches The manual annotation of AP2/ERF family from Eucalyptus grandis, Malus, Populus and Vitis genomes allowed a complete phylogenetic study for comparing the structure of this family in woody species and the model Arabidopsis thaliana. Expression profiles of the whole groups of EgrDREB1 and EgrDREB2 were investigated through RNAseq database survey and RT-qPCR analyses. Results The structure and the size of the AP2/ERF family show a global conservation for the plant species under comparison. In addition to an expansion of the ERF subfamily, the tree genomes mainly differ with respect to the group representation within the subfamilies. With regard to the E. grandis DREB subfamily, an obvious feature is the presence of 17 DREB1/CBF genes, the maximum reported to date for dicotyledons. In contrast, only six DREB2 have been identified, which is similar to the other plants species under study, except for Malus. All the DREB1/CBF and DREB2 genes from E. grandis are expressed in at least one condition and all are heat-responsive. Regulation by cold and drought depends on the genes but is not specific of one group; DREB1/CBF group is more cold-inducible than DREB2 which is mainly drought responsive. Conclusion These features suggest that the dramatic expansion of the DREB1/CBF group might be related to the adaptation of this evergreen tree to climate changes when it expanded in Australia. PMID:25849589

Genome-wide analysis of the AP2/ERF family in Eucalyptus grandis: an intriguing over-representation of stress-responsive DREB1/CBF genes.

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P B Cao

Full Text Available The AP2/ERF family includes a large number of developmentally and physiologically important transcription factors sharing an AP2 DNA-binding domain. Among them DREB1/CBF and DREB2 factors are known as master regulators respectively of cold and heat/osmotic stress responses.The manual annotation of AP2/ERF family from Eucalyptus grandis, Malus, Populus and Vitis genomes allowed a complete phylogenetic study for comparing the structure of this family in woody species and the model Arabidopsis thaliana. Expression profiles of the whole groups of EgrDREB1 and EgrDREB2 were investigated through RNAseq database survey and RT-qPCR analyses.The structure and the size of the AP2/ERF family show a global conservation for the plant species under comparison. In addition to an expansion of the ERF subfamily, the tree genomes mainly differ with respect to the group representation within the subfamilies. With regard to the E. grandis DREB subfamily, an obvious feature is the presence of 17 DREB1/CBF genes, the maximum reported to date for dicotyledons. In contrast, only six DREB2 have been identified, which is similar to the other plants species under study, except for Malus. All the DREB1/CBF and DREB2 genes from E. grandis are expressed in at least one condition and all are heat-responsive. Regulation by cold and drought depends on the genes but is not specific of one group; DREB1/CBF group is more cold-inducible than DREB2 which is mainly drought responsive.These features suggest that the dramatic expansion of the DREB1/CBF group might be related to the adaptation of this evergreen tree to climate changes when it expanded in Australia.

Molecular characterization of MHC-DRB cDNA in water buffalo (Bubalus bubalis

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Soumen Naskar

2012-01-01

Full Text Available In the present study, water buffalo MHC (Bubu-DRB cDNA was cloned and characterized. The 1022 base long-amplified cDNA product encompassed a single open reading frame of 801 bases that coded for 266 amino acids. The Bubu-DRB sequence showed maximum homology with the BoLA-DRB3*0101 allele of cattle. A total of seven amino acid residues were found to be unique for the Bubu-DRB sequence. The majority of amino acid substitutions was observed in the β1 domain. Residues associated with important functions were mostly conserved. Water buffalo DRB was phylogenetically closer to goat DRB*A.

Cost-effective sequencing of full-length cDNA clones powered by a de novo-reference hybrid assembly.

Science.gov (United States)

Kuroshu, Reginaldo M; Watanabe, Junichi; Sugano, Sumio; Morishita, Shinichi; Suzuki, Yutaka; Kasahara, Masahiro

2010-05-07

Sequencing full-length cDNA clones is important to determine gene structures including alternative splice forms, and provides valuable resources for experimental analyses to reveal the biological functions of coded proteins. However, previous approaches for sequencing cDNA clones were expensive or time-consuming, and therefore, a fast and efficient sequencing approach was demanded. We developed a program, MuSICA 2, that assembles millions of short (36-nucleotide) reads collected from a single flow cell lane of Illumina Genome Analyzer to shotgun-sequence approximately 800 human full-length cDNA clones. MuSICA 2 performs a hybrid assembly in which an external de novo assembler is run first and the result is then improved by reference alignment of shotgun reads. We compared the MuSICA 2 assembly with 200 pooled full-length cDNA clones finished independently by the conventional primer-walking using Sanger sequencers. The exon-intron structure of the coding sequence was correct for more than 95% of the clones with coding sequence annotation when we excluded cDNA clones insufficiently represented in the shotgun library due to PCR failure (42 out of 200 clones excluded), and the nucleotide-level accuracy of coding sequences of those correct clones was over 99.99%. We also applied MuSICA 2 to full-length cDNA clones from Toxoplasma gondii, to confirm that its ability was competent even for non-human species. The entire sequencing and shotgun assembly takes less than 1 week and the consumables cost only approximately US$3 per clone, demonstrating a significant advantage over previous approaches.

Extraction of Total DNA and RNA from Marine Filter Samples and Generation of a cDNA as Universal Template for Marker Gene Studies.

Science.gov (United States)

Schneider, Dominik; Wemheuer, Franziska; Pfeiffer, Birgit; Wemheuer, Bernd

2017-01-01

Microbial communities play an important role in marine ecosystem processes. Although the number of studies targeting marker genes such as the 16S rRNA gene has been increased in the last few years, the vast majority of marine diversity is rather unexplored. Moreover, most studies focused on the entire bacterial community and thus disregarded active microbial community players. Here, we describe a detailed protocol for the simultaneous extraction of DNA and RNA from marine water samples and for the generation of cDNA from the isolated RNA which can be used as a universal template in various marker gene studies.

Effects of Urban Waste and Mineral Fertilizers Applications on Eucalyptus grandis Growth and Soil Conditions Efeito da Aplicação de Lixo Urbano Compostado e de Adubos Minerais no Solo e na Produtividade de Eucalyptus grandis

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Guilherme de Castro Andrade

2011-03-01

Full Text Available

Effect of Thermo-Treatment on the Physical and Mechanical, Color, Fungal Durability of Wood of Tectona Grandis and Gmelina Arborea from Forest Plantations

Directory of Open Access Journals (Sweden)

Luis Diego MÉNDEZ-MEJÍAS

2018-02-01

Full Text Available This study evaluated the effect of thermo-treatment (THT at 4 temperatures on the density, shrinking, mass loss, moisture absorption, color, durability in terms of resistance to decay, flexural strength, tensile adhesion of glue line and the infrared spectrum of the wood of Tectona grandis and Gmelina arborea. Sapwood, heartwood and radial and tangential grain patterns were studied. The results showed that the THT temperature decreases the density, the percentage of moisture absorption, the modulus of elasticity and modulus of rupture in the flexure test and the tensile adhesion of glue line. The percentage of shrinking and durability presented irregular behavior relative to the THT temperature. The percentage of mass loss increased with increasing THT temperature in both species. The total color change (∆E* of thermo-treated wood (THTwood also increased with increasing THT temperature. Sapwood of T. grandis and G. arborea, having clearer shades, showed a more noticeable color change compared to hardwood; however, no significant differences were obtained between some of the THT temperatures.DOI: http://dx.doi.org/10.5755/j01.ms.24.1.17545

Cloning and functional expression of a cDNA encoding stearoyl-ACP Δ9-desaturase from the endosperm of coconut (Cocos nucifera L.).

Science.gov (United States)

Gao, Lingchao; Sun, Ruhao; Liang, Yuanxue; Zhang, Mengdan; Zheng, Yusheng; Li, Dongdong

2014-10-01

Coconut (Cocos nucifera L.) is an economically tropical fruit tree with special fatty acid compositions. The stearoyl-acyl carrier protein (ACP) desaturase (SAD) plays a key role in the properties of the majority of cellular glycerolipids. In this paper, a full-length cDNA of a stearoyl-acyl carrier protein desaturase, designated CocoFAD, was isolated from cDNA library prepared from the endosperm of coconut (C. nucifera L.). An 1176 bp cDNA from overlapped PCR products containing ORF encoding a 391-amino acid (aa) protein was obtained. The coded protein was virtually identical and shared the homology to other Δ9-desaturase plant sequences (greater than 80% as similarity to that of Elaeis guineensis Jacq). The real-time fluorescent quantitative PCR result indicated that the yield of CocoFAD was the highest in the endosperm of 8-month-old coconut and leaf, and the yield was reduced to 50% of the highest level in the endosperm of 15-month-old coconut. The coding region showed heterologous expression in strain INVSc1 of yeast (Saccharomyces cerevisiae). GC-MS analysis showed that the levels of palmitoleic acid (16:1) and oleic acid (18:1) were improved significantly; meanwhile stearic acid (18:0) was reduced. These results indicated that the plastidial Δ9 desaturase from the endosperm of coconut was involved in the biosynthesis of hexadecenoic acid and octadecenoic acid, which was similar with other plants. These results may be valuable for understanding the mechanism of fatty acid metabolism and the genetic improvement of CocoFAD gene in palm plants in the future. Copyright © 2014 Elsevier B.V. All rights reserved.

MOLECULAR CHARACTERIZATION OF ENDOCRINE DISRUPTION IN FISH USING CDNA ARRAYS.

Science.gov (United States)

We are developing cDNA macroarrays to measure the induction of gene expression in sheepshead minnows and largemouth bass exposed to anthropogenic chemicals that can mimic the action of endogenous hormones. For sheepshead minnows exposed in aqua, we observed similar genetic profil...

Transcription analysis of apple fruit development using cDNA microarrays

NARCIS (Netherlands)

Soglio, V.; Costa, F.; Molthoff, J.W.; Weemen-Hendriks, M.; Schouten, H.J.; Gianfranceschi, L.

2009-01-01

The knowledge of the molecular mechanisms underlying fruit quality traits is fundamental to devise efficient marker-assisted selection strategies and to improve apple breeding. In this study, cDNA microarray technology was used to identify genes whose expression changes during fruit development and

Isolation and molecular characterization of cathepsin L-like cysteine protease cDNAs from Western flower thrips (Frankliniella occidentalis)

NARCIS (Netherlands)

Kuipers, A.G.J.; Jongsma, M.A.

2004-01-01

Cysteine proteases are predominant in thrips guts (TGs) and, therefore, a suitable target for selecting effective protease inhibitors against western flower thrips (Frankliniella occidentalis). We report the isolation of four full-length cysteine protease cDNA clones from thrips in a two-step PCR

A new set of ESTs and cDNA clones from full-length and normalized libraries for gene discovery and functional characterization in citrus

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Alamar Santiago

2009-09-01

Full Text Available Abstract Background Interpretation of ever-increasing raw sequence information generated by modern genome sequencing technologies faces multiple challenges, such as gene function analysis and genome annotation. Indeed, nearly 40% of genes in plants encode proteins of unknown function. Functional characterization of these genes is one of the main challenges in modern biology. In this regard, the availability of full-length cDNA clones may fill in the gap created between sequence information and biological knowledge. Full-length cDNA clones facilitate functional analysis of the corresponding genes enabling manipulation of their expression in heterologous systems and the generation of a variety of tagged versions of the native protein. In addition, the development of full-length cDNA sequences has the power to improve the quality of genome annotation. Results We developed an integrated method to generate a new normalized EST collection enriched in full-length and rare transcripts of different citrus species from multiple tissues and developmental stages. We constructed a total of 15 cDNA libraries, from which we isolated 10,898 high-quality ESTs representing 6142 different genes. Percentages of redundancy and proportion of full-length clones range from 8 to 33, and 67 to 85, respectively, indicating good efficiency of the approach employed. The new EST collection adds 2113 new citrus ESTs, representing 1831 unigenes, to the collection of citrus genes available in the public databases. To facilitate functional analysis, cDNAs were introduced in a Gateway-based cloning vector for high-throughput functional analysis of genes in planta. Herein, we describe the technical methods used in the library construction, sequence analysis of clones and the overexpression of CitrSEP, a citrus homolog to the Arabidopsis SEP3 gene, in Arabidopsis as an example of a practical application of the engineered Gateway vector for functional analysis. Conclusion The new

A new set of ESTs and cDNA clones from full-length and normalized libraries for gene discovery and functional characterization in citrus

Science.gov (United States)

Marques, M Carmen; Alonso-Cantabrana, Hugo; Forment, Javier; Arribas, Raquel; Alamar, Santiago; Conejero, Vicente; Perez-Amador, Miguel A

2009-01-01

Background Interpretation of ever-increasing raw sequence information generated by modern genome sequencing technologies faces multiple challenges, such as gene function analysis and genome annotation. Indeed, nearly 40% of genes in plants encode proteins of unknown function. Functional characterization of these genes is one of the main challenges in modern biology. In this regard, the availability of full-length cDNA clones may fill in the gap created between sequence information and biological knowledge. Full-length cDNA clones facilitate functional analysis of the corresponding genes enabling manipulation of their expression in heterologous systems and the generation of a variety of tagged versions of the native protein. In addition, the development of full-length cDNA sequences has the power to improve the quality of genome annotation. Results We developed an integrated method to generate a new normalized EST collection enriched in full-length and rare transcripts of different citrus species from multiple tissues and developmental stages. We constructed a total of 15 cDNA libraries, from which we isolated 10,898 high-quality ESTs representing 6142 different genes. Percentages of redundancy and proportion of full-length clones range from 8 to 33, and 67 to 85, respectively, indicating good efficiency of the approach employed. The new EST collection adds 2113 new citrus ESTs, representing 1831 unigenes, to the collection of citrus genes available in the public databases. To facilitate functional analysis, cDNAs were introduced in a Gateway-based cloning vector for high-throughput functional analysis of genes in planta. Herein, we describe the technical methods used in the library construction, sequence analysis of clones and the overexpression of CitrSEP, a citrus homolog to the Arabidopsis SEP3 gene, in Arabidopsis as an example of a practical application of the engineered Gateway vector for functional analysis. Conclusion The new EST collection denotes an

阴道毛滴虫Rac1蛋白的cDNA克隆和序列分析%Molecular Cloning and Characterization of a Rac1 Homologue cDNA from Trichomonas vaginalis

Institute of Scientific and Technical Information of China (English)

傅玉才; 章家新; 郑晓虹; 刘红

2004-01-01

Objective To clone and characterize a Racl homologue from Trichomonas vaginalis for studying cell cycle of the organism. Methods A cDNA library derived from T. vaginalis mRNA was constructed into λ TriplEx2 phage vector. An expression sequence tag program was launched. Sequences of cDNA clones were analyzed using NCBI BLAST algorithms, and ClustalW and Treeview programs. Results A cDNA clone with a length of 714 base pairs was isolated. The sequence analysis showed that the cDNA clone has an open reading frame with 600 bp. The deduced amino acid sequence from the open reading frame contains 200 residuals and is most homologous to Rac1 subfamily of Rho GTPases with ＞ 60% identity. The conserved sequence elements of Rho GTPases, such as GTP-binding sites, GTPase-activating protein (GAP) interaction motifs, GTP-dissociation inhibitors (GDI) interaction motifs, guanine nucleotide exchange factor (GEF) interaction elements, etc, were detected in the amino acid sequence. The phylogenetic analysis showed that the cDNA clone is grouped in the Rac subfamily and is more closely related to Rac1 proteins of protozoa. Conclusion The cDNA clone isolated belongs to Rac subfamily of Rho GTPases and is probably a Rac1 protein of T. vaginalis.%目的获得阴道毛滴虫Rac1蛋白的cDNA克隆,研究其在细胞周期中的调解作用.方法提取阴道毛滴虫总RNA,构建cDNA表达文库,随机分离cDNA克隆并测序.用在线生物分析软件NCBI BLAST、ClustalW以及Treeview等程序进行序列分析.结果获得一株有714 bp的cDNA克隆.序列分析表明,该克隆开放阅读框具600 bp,推测肽链具200个氨基酸.该肽链与Rho家族中Rac1鸟苷三磷酸(GTP)酶同源性最高(＞60%),并具多种Rho GTP酶的保守基序,如GTP结合部位、GTP酶激活蛋白作用基序、GTP分离抑制因子作用基序、鸟嘌呤核苷酸交换因子作用基序等.进化树分析显示该克隆属于Rac亚家族GTP酶,与原虫Rac1蛋白最接近.结论该克隆�

The Eucalyptus grandis R2R3-MYB transcription factor family: evidence for woody growth-related evolution and function.

Science.gov (United States)

Soler, Marçal; Camargo, Eduardo Leal Oliveira; Carocha, Victor; Cassan-Wang, Hua; San Clemente, Hélène; Savelli, Bruno; Hefer, Charles A; Paiva, Jorge A Pinto; Myburg, Alexander A; Grima-Pettenati, Jacqueline

2015-06-01

The R2R3-MYB family, one of the largest transcription factor families in higher plants, controls a wide variety of plant-specific processes including, notably, phenylpropanoid metabolism and secondary cell wall formation. We performed a genome-wide analysis of this superfamily in Eucalyptus, one of the most planted hardwood trees world-wide. A total of 141 predicted R2R3-MYB sequences identified in the Eucalyptus grandis genome sequence were subjected to comparative phylogenetic analyses with Arabidopsis thaliana, Oryza sativa, Populus trichocarpa and Vitis vinifera. We analysed features such as gene structure, conserved motifs and genome location. Transcript abundance patterns were assessed by RNAseq and validated by high-throughput quantitative PCR. We found some R2R3-MYB subgroups with expanded membership in E. grandis, V. vinifera and P. trichocarpa, and others preferentially found in woody species, suggesting diversification of specific functions in woody plants. By contrast, subgroups containing key genes regulating lignin biosynthesis and secondary cell wall formation are more conserved across all of the species analysed. In Eucalyptus, R2R3-MYB tandem gene duplications seem to disproportionately affect woody-preferential and woody-expanded subgroups. Interestingly, some of the genes belonging to woody-preferential subgroups show higher expression in the cambial region, suggesting a putative role in the regulation of secondary growth. © 2014 The Authors New Phytologist © 2014 New Phytologist Trust.

Microbiome analysis shows enrichment for specific bacteria in separate anatomical regions of the deep-sea carnivorous sponge Chondrocladia grandis.

Science.gov (United States)

Verhoeven, Joost T P; Kavanagh, Alana N; Dufour, Suzanne C

2017-01-01

The Cladorhizidae is a unique family of carnivorous marine sponges characterised by either the absence or reduction of the aquiferous system and by the presence of specialised structures to trap and digest mesoplanktonic prey. Previous studies have postulated a key role of host-associated bacteria in enabling carnivory in this family of sponges. In this study, we employed high-throughput Illumina-based sequencing to identify the bacterial community associated with four individuals of the deep-sea sponge Chondrocladia grandis sampled in the Gulf of Maine. By characterising the V6 through V8 region of the 16S rRNA gene, we compared the bacterial community composition and diversity in three distinct anatomical regions with predicted involvement in prey capture (sphere), support (axis) and benthic substrate attachment (root). A high abundance of Tenacibaculum, a known siderophore producing bacterial genus, was present in all anatomical regions and specimens. The abundance of Colwellia and Roseobacter was greater in sphere and axis samples, and bacteria from the hydrocarbon-degrading Robiginitomaculum genus were most abundant in the root. This first description of the bacterial community associated with C. grandis provides novel insights into the contribution of bacteria to the carnivorous lifestyle while laying foundations for future cladorhizid symbiosis studies. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Generation of an infectious clone of a new Korean isolate of apple chlorotic leaf spot virus (ACLSV) driven by dual 35S and T7 promoters in a versatile binary vector

Science.gov (United States)

The full-length sequence of a new isolate of Apple chlorotic leaf spot virus (ACLSV) from Korea was divergent, but most closely related to the Japanese isolate A4, at 84% nucleotide identity. The full-length cDNA of the Korean isolate of ACLSV was cloned into a binary vector downstream of the bacter...

FIELD VALIDATION OF A SHEEPSHEAD MINNOW ESTROGEN-RESPONSIVE CDNA MACROARRAY

Science.gov (United States)

Hemmer, Michael J., Iris Knoebl, Becky L. Hemmer, Patrick Larkin, Peggy S. Harris and Nancy D. Denslow. In press. Field Validation of a Sheepshead Minnow Estrogen-Responsive cDNA Macroarray (Abstract). To be presented at the SETAC Fourth World Congress, 14-18 November 2004, Portl...

Características tecnológicas da madeira de árvores matrizes de Eucalyptus grandis.

Directory of Open Access Journals (Sweden)

Clovis Roberto Haselein

2010-08-01

Full Text Available O objetivo do presente trabalho foi caracterizar árvores matrizes de Eucalyptus grandis em relação a características tecnológicas da madeira. Foram selecionadas 63 árvores matrizes, pertencentes a um povoamento comercial localizado no litoral norte do Rio Grande do Sul. As árvores foram abatidas e, com base nos dados dendrométricos, calculou-se o volume comercial com e sem casca, o fator de forma, a conicidade e a relação altura/diâmetro. Quanto à madeira, foi analisada a massa específica básica, os percentuais volumétricos de cerne, alburno e casca, as rachaduras de topo das toras e das tábuas, os empenamentos e os defeitos visuais das tábuas, tais como nós e bolsa de resina. A massa específica básica, cuja média inclui a madeira dessa espécie entre as moderadamente leves a pesadas, mostrou uma tendência decrescente entre o DAP e 25% da altura comercial e, a partir daí, crescente até 100% da mesma. O percentual volumétrico de cerne apresentou valores médios crescentes desde a base até 25% da altura comercial e, a partir daí, diminuiu até 100% da altura comercial, sendo que a média geral foi de 75,7%. O comprimento médio das rachaduras de topo em tábuas de Eucalyptus grandis foi de 63%. A alta porcentagem de rachaduras de topo foi atribuída à posição de retirada das tábuas, próxima à medula. Já a variabilidade das rachaduras de topo encontrada para as toras (CV=60% possibilita a seleção de árvores com menor tendência em apresentar esse defeito.

Temperature effects on wood anatomy, wood density, photosynthesis and biomass partitioning of Eucalyptus grandis seedlings.

Science.gov (United States)

Thomas, D S; Montagu, K D; Conroy, J P

2007-02-01

Wood density, a gross measure of wood mass relative to wood volume, is important in our understanding of stem volume growth, carbon sequestration and leaf water supply. Disproportionate changes in the ratio of wood mass to volume may occur at the level of the whole stem or the individual cell. In general, there is a positive relationship between temperature and wood density of eucalypts, although this relationship has broken down in recent years with wood density decreasing as global temperatures have risen. To determine the anatomical causes of the effects of temperature on wood density, Eucalyptus grandis W. Hill ex Maiden seedlings were grown in controlled-environment cabinets at constant temperatures from 10 to 35 degrees C. The 20% increase in wood density of E. grandis seedlings grown at the higher temperatures was variously related to a 40% reduction in lumen area of xylem vessels, a 10% reduction in the lumen area of fiber cells and a 10% increase in fiber cell wall thickness. The changes in cell wall characteristics could be considered analogous to changes in carbon supply. Lumen area of fiber cells declined because of reduced fiber cell expansion and increased fiber cell wall thickening. Fiber cell wall thickness was positively related to canopy CO2 assimilation rate (Ac), which increased 26-fold because of a 24-fold increase in leaf area and a doubling in leaf CO2 assimilation rate from minima at 10 and 35 degrees C to maxima at 25 and 30 degrees C. Increased Ac increased seedling volume, biomass and wood density; but increased wood density was also related to a shift in partitioning of seedling biomass from roots to stems as temperature increased.

Isolation and Characterization of D-Myo-Inositol-3-Phosphate Synthase Gene Family Members in Soybean

OpenAIRE

Good, Laura Lee

2001-01-01

The objective of this research was to isolate genes encoding isoforms of the enzyme D-myo-inositol 3-phosphate synthase (MIPS, E.C. 5.5.1.4) from soybean and to characterize their expression, especially with respect to their involvement in phytic acid biosynthesis. A MIPS-homologous cDNA, designated GmMIPS1, was isolated via PCR using total RNA from developing seeds. Southern blot analysis and examination of MIPS-homologous soybean EST sequences suggested that GmMIPS1 is part of a multigene...

cDNA, genomic sequence cloning and overexpression of ribosomal ...

African Journals Online (AJOL)

RPS16 of eukaryote is a component of the 40S small ribosomal subunit encoded by RPS16 gene and is also a homolog of prokaryotic RPS9. The cDNA and genomic sequence of RPS16 was cloned successfully for the first time from the Giant Panda (Ailuropoda melanoleuca) using reverse transcription-polymerase chain ...

Complete cDNA sequence coding for human docking protein

Energy Technology Data Exchange (ETDEWEB)

Hortsch, M; Labeit, S; Meyer, D I

1988-01-11

Docking protein (DP, or SRP receptor) is a rough endoplasmic reticulum (ER)-associated protein essential for the targeting and translocation of nascent polypeptides across this membrane. It specifically interacts with a cytoplasmic ribonucleoprotein complex, the signal recognition particle (SRP). The nucleotide sequence of cDNA encoding the entire human DP and its deduced amino acid sequence are given.

Cloning and characterization of transferrin cDNA and rapid detection of transferrin gene polymorphism in rainbow trout (Oncorhynchus mykiss).

Science.gov (United States)

Tange, N; Jong-Young, L; Mikawa, N; Hirono, I; Aoki, T

1997-12-01

A cDNA clone of rainbow trout (Oncorhynchus mykiss) transferrin was obtained from a liver cDNA library. The 2537-bp cDNA sequence contained an open reading frame encoding 691 amino acids and the 5' and 3' noncoding regions. The amino acid sequences at the iron-binding sites and the two N-linked glycosylation sites, and the cysteine residues were consistent with known, conserved vertebrate transferrin cDNA sequences. Single N-linked glycosylation sites existed on the N- and C-lobe. The deduced amino acid sequence of the rainbow trout transferrin cDNA had 92.9% identities with transferrin of coho salmon (Oncorhynchus kisutch); 85%, Atlantic salmon (Salmo salar); 67.3%, medaka (Oryzias latipes); 61.3% Atlantic cod (Gadus morhua); and 59.7%, Japanese flounder (Paralichthys olivaceus). The long and accurate polymerase chain reaction (LA-PCR) was used to amplify approximately 6.5 kb of the transferrin gene from rainbow trout genomic DNA. Restriction fragment length polymorphisms (RFLPs) of the LA-PCR products revealed three digestion patterns in 22 samples.

Fiscal 1998 achievement report. Industrial technology research and development project. (Strategic human cDNA genome application technology development); 1998 nendo senryakuteki hito cDNA genome oyo gijutsu kaihatsu seika hokokusho

Energy Technology Data Exchange (ETDEWEB)

NONE

2000-03-01

A human genome related project named above was started, and studies were conducted for base sequence determination and function analysis for approximately 10,000 kinds of full-length or long-chain human cDNA clones owned by research organizations in this country. The Institute of Medical Science of University of Tokyo and Helix Research Institute dealt with a full-length human cDNA library constructed by oligo-capping, and determined the base sequences of all specimens in the library. The Kazusa DNA Research Institute determined partial sequences for long-chain clones which are not shorter than 4-5kbp, and determined entire sequences for some bases. The obtained base sequence data were subjected to homology analysis, the base sequences were converted into amino acid sequences, and functions of proteins were predicted. In the analysis of gene functions, ATAC-PCR (adaptor tagged competitive-polymerase chain reaction) was applied to the clones covered by this project, and a database was prepared by use of the results of analyses of frequency-related information. For the preparation of a comprehensive gene expression profile, technologies for cDNA microarray construction were established. (NEDO)

Microdissection and molecular manipulation of single chromosomes in woody fruit trees with small chromosomes using pomelo (Citrus grandis) as a model. I. Construction of single chromosomal DNA libraries.

Science.gov (United States)

Huang, D; Wu, W; Zhou, Y; Hu, Z; Lu, L

2004-05-01

Construction of single chromosomal DNA libraries by means of chromosome microdissection and microcloning will be useful for genomic research, especially for those species that have not been extensively studied genetically. Application of the technology of microdissection and microcloning to woody fruit plants has not been reported hitherto, largely due to the generally small sizes of metaphase chromosomes and the difficulty of chromosome preparation. The present study was performed to establish a method for single chromosome microdissection and microcloning in woody fruit species using pomelo as a model. The standard karyotype of a pomelo cultivar ( Citrus grandis cv. Guanxi) was established based on 20 prometaphase photomicrographs. According to the standard karyotype, chromosome 1 was identified and isolated with fine glass microneedles controlled by a micromanipulator. DNA fragments ranging from 0.3 kb to 2 kb were acquired from the isolated single chromosome 1 via two rounds of PCR mediated by Sau3A linker adaptors and then cloned into T-easy vectors to generate a DNA library of chromosome 1. Approximately 30,000 recombinant clones were obtained. Evaluation based on 108 randomly selected clones showed that the sizes of the cloned inserts varied from 0.5 kb to 1.5 kb with an average of 860 bp. Our research suggests that microdissection and microcloning of single small chromosomes in woody plants is feasible.

Phytochemical profile, aldose reductase inhibitory, and antioxidant activities of Indian traditional medicinal Coccinia grandis (L.) fruit extract.

Science.gov (United States)

Kondhare, Dasharath; Lade, Harshad

2017-12-01

Coccinia grandis (L.) fruits (CGFs) are commonly used for culinary purposes and has several therapeutic applications in the Southeast Asia. The aim of this work was to evaluate phytochemical profile, aldose reductase inhibitory (ARI), and antioxidant activities of CGF extract. The CGFs were extracted with different solvents including petroleum ether, dichloromethane, acetone, methanol, and water. The highest yield of total extractable compounds (34.82%) and phenolic content (11.7 ± 0.43 mg of GAE/g dried extract) was found in methanol extract, whereas water extract showed the maximum content of total flavonoids (82.8 ± 7.8 mg QE/g dried extract). Gas chromatography-mass spectroscopy (GC-MS) analysis of methanol and water extract revealed the presence of flavonoids, phenolic compounds, alkaloids, and glycosides in the CGFs. Results of the in vitro ARI activity against partially purified bovine lens aldose reductase showed that methanol extract of CGFs exhibited 96.6% ARI activity at IC 50 value 6.12 µg/mL followed by water extract 89.1% with the IC 50 value 6.50 µg/mL. In addition, methanol and water extracts of CGF showed strong antioxidant activities including ABTS *+ scavenging, DPPH* scavenging, and hydroxyl radical scavenging. Our results suggest that high percentage of both flavonoids and phenolic contents in the CGFs are correlated with the ARI and antioxidant activities. The fruits of C. grandis are thus potential bifunctional agents with ARI and antioxidant activities that can be used for the prevention and management of DM and associated diseases.

cDNA sequences of two apolipoproteins from lamprey

International Nuclear Information System (INIS)

Pontes, M.; Xu, X.; Graham, D.; Riley, M.; Doolittle, R.F.

1987-01-01

The messages for two small but abundant apolipoproteins found in lamprey blood plasma were cloned with the aid of oligonucleotide probes based on amino-terminal sequences. In both cases, numerous clones were identified in a lamprey liver cDNA library, consistent with the great abundance of these proteins in lamprey blood. One of the cDNAs (LAL1) has a coding region of 105 amino acids that corresponds to a 21-residue signal peptide, a putative 8-residue propeptide, and the 76-residue mature protein found in blood. The other cDNA (LAL2) codes for a total of 191 residues, the first 23 of which constitute a signal peptide. The two proteins, which occur in the high-density lipoprotein fraction of ultracentrifuged plasma, have amino acid compositions similar to those of apolipoproteins found in mammalian blood; computer analysis indicates that the sequences are largely helix-permissive. When the sequences were searched against an amino acid sequence data base, rat apolipoprotein IV was the best matching candidate in both cases. Although a reasonable alignment can be made with that sequence and LAL1, definitive assignment of the two lamprey proteins to typical mammalian classes cannot be made at this point

Rat organic solute carrier protein 1 (rOscp1) mediated the transport of organic solutes in Xenopus laevis oocytes: isolation and pharmacological characterization of rOscp1.

Science.gov (United States)

Izuno, Hisanori; Kobayashi, Yasuna; Sanada, Yutaka; Nihei, Daisuke; Suzuki, Masako; Kohyama, Noriko; Ohbayashi, Masayuki; Yamamoto, Toshinori

2007-09-22

Rat organic solute carrier protein 1 (rOscp1) was isolated from a rat testis cDNA library. Isolated rOscp1 cDNA consisted of 1089 base pairs that encoded a 363-amino acid protein, and the amino acid sequence was 88% and 93% identical to that of human OSCP1 (hOSCP1) and mouse Oscp1 (mOscp1), respectively. The message for rOscp1 is highly detected in rat testis. When expressed in X. oocytes, rOscp1 mediated the high affinity transport of p-aminohippurate (PAH) with a Km value of 15.7+/-1.9 microM, and rOscp1-mediated organic solutes were exhibited in time- and Na+-independent manners. rOscp1 also transported various structurally heterogenous compounds such as testosterone, dehydroepiandrosterone sulfate (DHEA-S), and taurocholate with some differences in substrate specificity compared with hOSCP1. Immunohistochemical analysis revealed that the rOscp1 protein is localized in the basal membrane side of Sertoli cells as observed in mouse testis [Kobayashi et al., 2007; Kobayashi, Y., Tsuchiya, A., Hayashi, T., Kohyama, N., Ohbayashi, M., Yamamoto, T., 2007. Isolation and characterization of polyspecific mouse organic solute carrier protein 1 (mOscp1). Drug Metabolism and Disposition 35 (7), 1239-1245]. Thus, the present results indicate that a newly isolated cDNA clone, rOscp1, is a polyspecific organic solute carrier protein with some differences in substrate specificity compared with human and mouse OSCP1.

Benchmarking of the Oxford Nanopore MinION sequencing for quantitative and qualitative assessment of cDNA populations.

Science.gov (United States)

Oikonomopoulos, Spyros; Wang, Yu Chang; Djambazian, Haig; Badescu, Dunarel; Ragoussis, Jiannis

2016-08-24

To assess the performance of the Oxford Nanopore Technologies MinION sequencing platform, cDNAs from the External RNA Controls Consortium (ERCC) RNA Spike-In mix were sequenced. This mix mimics mammalian mRNA species and consists of 92 polyadenylated transcripts with known concentration. cDNA libraries were generated using a template switching protocol to facilitate the direct comparison between different sequencing platforms. The MinION performance was assessed for its ability to sequence the cDNAs directly with good accuracy in terms of abundance and full length. The abundance of the ERCC cDNA molecules sequenced by MinION agreed with their expected concentration. No length or GC content bias was observed. The majority of cDNAs were sequenced as full length. Additionally, a complex cDNA population derived from a human HEK-293 cell line was sequenced on an Illumina HiSeq 2500, PacBio RS II and ONT MinION platforms. We observed that there was a good agreement in the measured cDNA abundance between PacBio RS II and ONT MinION (rpearson = 0.82, isoforms with length more than 700bp) and between Illumina HiSeq 2500 and ONT MinION (rpearson = 0.75). This indicates that the ONT MinION can sequence quantitatively both long and short full length cDNA molecules.

A Full-Length Infectious cDNA Clone of Zika Virus from the 2015 Epidemic in Brazil as a Genetic Platform for Studies of Virus-Host Interactions and Vaccine Development

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Konstantin A. Tsetsarkin

2016-08-01

Full Text Available An arthropod-borne virus, Zika virus (ZIKV, has recently emerged as a major human pathogen. Associated with complications during perinatal development and Guillain-Barré syndrome in adults, ZIKV raises new challenges for understanding the molecular determinants of flavivirus pathogenesis. This underscores the necessity for the development of a reverse genetic system based on an epidemic ZIKV strain. Here, we describe the generation and characterization in cell cultures of an infectious cDNA clone of ZIKV isolated from the 2015 epidemic in Brazil. The cDNA-derived ZIKV replicated efficiently in a variety of cell lines, including those of both neuronal and placental origin. We observed that the growth of cDNA-derived virus was attenuated compared to the growth of the parental isolate in most cell lines, which correlates with substantial differences in sequence heterogeneity between these viruses that were determined by deep-sequencing analysis. Our findings support the role of genetic diversity in maintaining the replicative fitness of viral populations under changing conditions. Moreover, these results indicate that caution should be exercised when interpreting the results of reverse-genetics experiments in attempts to accurately predict the biology of natural viruses. Finally, a Vero cell-adapted cDNA clone of ZIKV was generated that can be used as a convenient platform for studies aimed at the development of ZIKV vaccines and therapeutics.

Application of a cDNA microarray for profiling the gene expression of Echinococcus granulosus protoscoleces treated with albendazole and artemisinin.

Science.gov (United States)

Lü, Guodong; Zhang, Wenbao; Wang, Jianhua; Xiao, Yunfeng; Zhao, Jun; Zhao, Jianqin; Sun, Yimin; Zhang, Chuanshan; Wang, Junhua; Lin, Renyong; Liu, Hui; Zhang, Fuchun; Wen, Hao

2014-12-01

Cystic echinoccocosis (CE) is a neglected zoonosis that is caused by the dog-tapeworm Echinococcus granulosus. The disease is endemic worldwide. There is an urgent need for searching effective drug for the treatment of the disease. In this study, we sequenced a cDNA library constructed using RNA isolated from oncospheres, protoscoleces, cyst membrane and adult worms of E. granulosus. A total of 9065 non-redundant or unique sequences were obtained and spotted on chips as uniEST probes to profile the gene expression in protoscoleces of E. granulosus treated with the anthelmintic drugs albendazole and artemisinin, respectively. The results showed that 7 genes were up-regulated and 38 genes were down-regulated in the protoscoleces treated with albendazole. Gene analysis showed that these genes are responsible for energy metabolism, cell cycle and assembly of cell structure. We also identified 100 genes up-regulated and 6 genes down-regulated in the protoscoleces treated with artemisinin. These genes play roles in the transduction of environmental signals, and metabolism. Albendazole appeared its drug efficacy in damaging cell structure, while artemisinin was observed to increase the formation of the heterochromatin in protoscolex cells. Our results highlight the utility of using cDNA microarray methods to detect gene expression profiles of E. granulosus and, in particular, to understand the pharmacologic mechanism of anti-echinococcosis drugs. Copyright © 2014 Elsevier B.V. All rights reserved.

A linear concatenation strategy to construct 5'-enriched amplified cDNA libraries using multiple displacement amplification.

Science.gov (United States)

Gadkar, Vijay J; Filion, Martin

2013-06-01

In various experimental systems, limiting available amounts of RNA may prevent a researcher from performing large-scale analyses of gene transcripts. One way to circumvent this is to 'pre-amplify' the starting RNA/cDNA, so that sufficient amounts are available for any downstream analysis. In the present study, we report the development of a novel protocol for constructing amplified cDNA libraries using the Phi29 DNA polymerase based multiple displacement amplification (MDA) system. Using as little as 200 ng of total RNA, we developed a linear concatenation strategy to make the single-stranded cDNA template amenable for MDA. The concatenation, made possible by the template switching property of the reverse transcriptase enzyme, resulted in the amplified cDNA library with intact 5' ends. MDA generated micrograms of template, allowing large-scale polymerase chain reaction analyses or other large-scale downstream applications. As the amplified cDNA library contains intact 5' ends, it is also compatible with 5' RACE analyses of specific gene transcripts. Empirical validation of this protocol is demonstrated on a highly characterized (tomato) and an uncharacterized (corn gromwell) experimental system.

Identification of a cryptic prokaryotic promoter within the cDNA encoding the 5' end of dengue virus RNA genome.

Directory of Open Access Journals (Sweden)

Dongsheng Li

Full Text Available Infectious cDNA clones of RNA viruses are important research tools, but flavivirus cDNA clones have proven difficult to assemble and propagate in bacteria. This has been attributed to genetic instability and/or host cell toxicity, however the mechanism leading to these difficulties has not been fully elucidated. Here we identify and characterize an efficient cryptic bacterial promoter in the cDNA encoding the dengue virus (DENV 5' UTR. Following cryptic transcription in E. coli, protein expression initiated at a conserved in-frame AUG that is downstream from the authentic DENV initiation codon, yielding a DENV polyprotein fragment that was truncated at the N-terminus. A more complete understanding of constitutive viral protein expression in E. coli might help explain the cloning and propagation difficulties generally observed with flavivirus cDNA.

Generation and analysis of large-scale expressed sequence tags (ESTs from a full-length enriched cDNA library of porcine backfat tissue

Directory of Open Access Journals (Sweden)

Lee Hae-Young

2006-02-01

Full Text Available Abstract Background Genome research in farm animals will expand our basic knowledge of the genetic control of complex traits, and the results will be applied in the livestock industry to improve meat quality and productivity, as well as to reduce the incidence of disease. A combination of quantitative trait locus mapping and microarray analysis is a useful approach to reduce the overall effort needed to identify genes associated with quantitative traits of interest. Results We constructed a full-length enriched cDNA library from porcine backfat tissue. The estimated average size of the cDNA inserts was 1.7 kb, and the cDNA fullness ratio was 70%. In total, we deposited 16,110 high-quality sequences in the dbEST division of GenBank (accession numbers: DT319652-DT335761. For all the expressed sequence tags (ESTs, approximately 10.9 Mb of porcine sequence were generated with an average length of 674 bp per EST (range: 200–952 bp. Clustering and assembly of these ESTs resulted in a total of 5,008 unique sequences with 1,776 contigs (35.46% and 3,232 singleton (65.54% ESTs. From a total of 5,008 unique sequences, 3,154 (62.98% were similar to other sequences, and 1,854 (37.02% were identified as having no hit or low identity (Sus scrofa. Gene ontology (GO annotation of unique sequences showed that approximately 31.7, 32.3, and 30.8% were assigned molecular function, biological process, and cellular component GO terms, respectively. A total of 1,854 putative novel transcripts resulted after comparison and filtering with the TIGR SsGI; these included a large percentage of singletons (80.64% and a small proportion of contigs (13.36%. Conclusion The sequence data generated in this study will provide valuable information for studying expression profiles using EST-based microarrays and assist in the condensation of current pig TCs into clusters representing longer stretches of cDNA sequences. The isolation of genes expressed in backfat tissue is the

Cloning and expression of a cDNA coding for a human monocyte-derived plasminogen activator inhibitor

International Nuclear Information System (INIS)

Antalis, T.M.; Clark, M.A.; Barnes, T.; Lehrbach, P.R.; Devine, P.L.; Schevzov, G.; Goss, N.H.; Stephens, R.W.; Tolstoshev, P.

1988-01-01

Human monocyte-derived plasminogen activator inhibitor (mPAI-2) was purified to homogeneity from the U937 cell line and partially sequenced. Oligonucleotide probes derived from this sequence were used to screen a cDNA library prepared from U937 cells. One positive clone was sequenced and contained most of the coding sequence as well as a long incomplete 3' untranslated region (1112 base pairs). This cDNA sequence was shown to encode mPAI-2 by hybrid-select translation. A cDNA clone encoding the remainder of the mPAI-2 mRNA was obtained by primer extension of U937 poly(A) + RNA using a probe complementary to the mPAI-2 coding region. The coding sequence for mPAI-2 was placed under the control of the λ P/sub L/ promoter, and the protein expressed in Escherichia coli formed a complex with urokinase that could be detected immunologically. By nucleotide sequence analysis, mPAI-2 cDNA encodes a protein containing 415 amino acids with a predicted unglycosylated M/sub r/ of 46,543. The predicted amino acid sequence of mPAI-2 is very similar to placental PAI-2 and shows extensive homology with members of the serine protease inhibitor (serpin) superfamily. mPAI-2 was found to be more homologous to ovalbumin (37%) than the endothelial plasminogen activator inhibitor, PAI-1 (26%). The 3' untranslated region of the mPAI-2 cDNA contains a putative regulatory sequence that has been associated with the inflammatory mediators

Construction of C35 gene bait recombinants and T47D cell cDNA library.

Science.gov (United States)

Yin, Kun; Xu, Chao; Zhao, Gui-Hua; Liu, Ye; Xiao, Ting; Zhu, Song; Yan, Ge

2017-11-20

C35 is a novel tumor biomarker associated with metastasis progression. To investigate the interaction factors of C35 in its high expressed breast cancer cell lines, we constructed bait recombinant plasmids of C35 gene and T47D cell cDNA library for yeast two-hybrid screening. Full length C35 sequences were subcloned using RT-PCR from cDNA template extracted from T47D cells. Based on functional domain analysis, the full-length C35 1-348bp was also truncated into two fragments C351-153bp and C35154-348bp to avoid auto-activation. The three kinds of C35 genes were successfully amplified and inserted into pGBKT7 to construct bait recombinant plasmids pGBKT7-C351-348bp, pGBKT7-C351-153bp and pGBKT7-C35154-348bp, then transformed into Y187 yeast cells by the lithium acetate method. Auto-activation and toxicity of C35 baits were detected using nutritional deficient medium and X-α-Gal assays. The T47D cell ds cDNA was generated by SMART TM technology and the library was constructed using in vivo recombination-mediated cloning in the AH109 yeast strain using a pGADT7-Rec plasmid. The transformed Y187/pGBKT7-C351-348bp line was intensively inhibited while the truncated Y187/pGBKT7-C35 lines had no auto-activation and toxicity in yeast cells. The titer of established cDNA library was 2 × 10 7 pfu/mL with high transformation efficiency of 1.4 × 10 6 , and the insert size of ds cDNA was distributed homogeneously between 0.5-2.0 kb. Our research generated a T47D cell cDNA library with high titer, and the constructed two C35 "baits" contained a respective functional immunoreceptor tyrosine based activation motif (ITAM) and the conserved last four amino acids Cys-Ile-Leu-Val (CILV) motif, and therefore laid a foundation for screening the C35 interaction factors in a BC cell line.

Detection of reverse transcriptase termination sites using cDNA ligation and massive parallel sequencing

DEFF Research Database (Denmark)

Kielpinski, Lukasz J; Boyd, Mette; Sandelin, Albin

2013-01-01

Detection of reverse transcriptase termination sites is important in many different applications, such as structural probing of RNAs, rapid amplification of cDNA 5' ends (5' RACE), cap analysis of gene expression, and detection of RNA modifications and protein-RNA cross-links. The throughput...... of these methods can be increased by applying massive parallel sequencing technologies.Here, we describe a versatile method for detection of reverse transcriptase termination sites based on ligation of an adapter to the 3' end of cDNA with bacteriophage TS2126 RNA ligase (CircLigase™). In the following PCR...

Isolation of oogenesis-specific genes transcribed in the germ-line of Calliphora erythrocephala and Drosophila melanogaster

International Nuclear Information System (INIS)

Tucker, M.A.

1988-01-01

Poly(A) + RNA from early or mid-stage ovarian follicles of C. erythrocephala was used to generate radiolabelled oogenesis-specific cDNA probes for screening the phage libraries. A cDNA probe made from mid-stage embryo poly(A) + RNA was used as the differential screening probe. Thus plaques hybridizing to the two oogenesis-specific probes but not the mid-stage embryo probe were selected as potentially containing oogenesis-specific genes. Two further rounds of screening were used to eliminate false positives and, after plaque purification, restriction digests of the remaining clones were screened by Southern blot hybridization to identify DNA fragments transcribed in an oogenesis-specific manner. In situ hybridization to sections of ovarian follicles has been used to determine the cell types within the follicles in which the various genes are expressed. Radiolabelled RNA probes for four of the C. erythrocephala oogenesis-specific clones and the two D. melanogaster clones have been hybridized to ovarian follicles. Further studies have been concentrated on the two germ-line transcribed, oogenesis-specific clones isolated from the D. melanogaster clone library. Detailed genetic mapping of the DA clone and of these mutations was performed to determine which mutations might represent the DA gene. cDNA clones have been isolated for the transcribed region of clone DA and have been used to further define the transcription unit from this region of the D. melanogaster genome

Enhanced specificity in immunoscreening of expression cDNA clones using radiolabeled antigen overlay

International Nuclear Information System (INIS)

Chao, S.; Chao, L.; Chao, J.

1989-01-01

A highly sensitive and specific method has been developed for immunoscreening clones from an expression cDNA library. The procedures utilize a radiolabeled antigen detection method described originally for the immunoblotting of plasma proteins. Screening of rat alpha 1-antitrypsin clones was used. Comparison between Western blots of alpha 1-antitrypsin using both labeled antigen and protein A detection methods showed that the former yielded lower background and greater sensitivity than the latter. Further, this technique was shown to have a lower detection limit of less than 20 ng through Western blot analysis of varying concentrations of alpha 1-antitrypsin. The procedures are based on the expression of the protein by cDNA clones containing the DNA inserts in the correct reading frame. Following the transfer of phage proteins to nitrocellulose membranes, the bivalent antibodies bind monovalently to both nitrocellulose-bound-antigen in the phage lysates and radiolabeled antigen. The radiolabeled antigen overlay method is superior to the protein A detection method in sensitivity, specificity and reproducibility. This improved method can be applied in general for screening expression cDNA libraries, provided that the specific antiserum and radiolabeled antigen are available

Isolation of a polyphenol oxidase (PPO) cDNA from artichoke and expression analysis in wounded artichoke heads.

Science.gov (United States)

Quarta, Angela; Mita, Giovanni; Durante, Miriana; Arlorio, Marco; De Paolis, Angelo

2013-07-01

The polyphenol oxidase (PPO) enzyme, which can catalyze the oxidation of phenolics to quinones, has been reported to be involved in undesirable browning in many plant foods. This phenomenon is particularly severe in artichoke heads wounded during the manufacturing process. A full-length cDNA encoding for a putative polyphenol oxidase (designated as CsPPO) along with a 1432 bp sequence upstream of the starting ATG codon was characterized for the first time from [Cynara cardunculus var. scolymus (L.) Fiori]. The 1764 bp CsPPO sequence encodes a putative protein of 587 amino acids with a calculated molecular mass of 65,327 Da and an isoelectric point of 5.50. Analysis of the promoter region revealed the presence of cis-acting elements, some of which are putatively involved in the response to light and wounds. Expression analysis of the gene in wounded capitula indicated that CsPPO was significantly induced after 48 h, even though the browning process had started earlier. This suggests that the early browning event observed in artichoke heads was not directly related to de novo mRNA synthesis. Finally, we provide the complete gene sequence encoding for polyphenol oxidase and the upstream regulative region in artichoke. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Qualidade de mudas clonais de Eucalyptus urophylla x E. grandis em função do substrato Quality of Eucalyptus urophylla x E. grandis cuttings as a function of substrate

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Richardson B. G. da Silva

2012-01-01

Full Text Available Considerando a crescente demanda por mudas florestais e a escassez de matérias-primas dos substratos utilizados para o crescimento das plantas, faz-se necessário avaliar novos componentes e formulações que assegurem a qualidade das mudas. Nesse experimento foram estudados o desenvolvimento e a qualidade de mudas clonais de Eucalyptus urophylla x E. grandis, provenientes de miniestacas, em função de nove composições de substrato produzidos a partir de vermiculita granulometria fina, casca de arroz carbonizada e fibra de coco em tubetes de 50 mL. Foram avaliados as propriedades físicas dos substratos, o desenvolvimento morfológico e a qualidade do sistema radicular das mudas, aos 90 dias após o estaqueamento. Os substratos com maior porosidade total promovem maior qualidade do sistema radicular o que, consequentemente, resulta em mudas com maior diâmetro, massa seca aérea e radicular e Índice de Qualidade de Dickson. Outros valores para as características físicas dos substratos, diferentes dos citados na literatura, também podem ser considerados adequados.Considering the increasing demand for seedlings of plants for aforestation and the scarcity of raw materials for the substrates used for plant growth, it is necessary to evaluate new components and formulations that ensure the quality of seedlings. In this experiment the development and quality of production of Eucalyptus urophylla x E. grandis seedlings was studied through vegetative propagation, as a function of nine compositions of substrate, produced from fine-grained vermiculite, carbonized rice chaff and coconut fiber in hard plastic tubes of 50 mL as containers. The physical properties of substrates and morphological development and the quality of the root system of seedlings were evaluated at 90 days after staking. The substrates with higher total porosity promotes better quality of the root system, which consequently results in cuttings with larger diameter, shoot and

Isolation, characterization, and mapping of gene encoding dihydrolipoyl succinyltransferase (E2k) of human [alpha]-ketoglutarate dehydrogenase complex

Energy Technology Data Exchange (ETDEWEB)

Ali, G.; Cai, Xingang; Sheu, Kwan-Fu R.; Blass, J.P. (Cornell Univ. Medical College, White Plains, NY (United States)); Wasco, W.; Gaston, S.M.; Tanzi, R.E.; Cooper, A.J.L.; Gusella, J.F. (Massachusetts General Hospital, Charleston, MA (United States)); Szabo, P. (Cornell Univ. Medical College, New York, NY (United States))

1994-03-01

The authors have isolated and sequenced cDNAs representing the full-length (2987-bp) gene for dihydrolipoyl succinyltransferase (E2k component) of the human [alpha]-ketoglutarate dehydrogenase complex (KHDHC) from a human fetal brain cDNA library. The E2k cDNA was mapped to human chromosome 14 using a somatic cell hybrid panel, and more precisely to band 14q24.3 by in situ hybridization. This cDNA also cross-hybridized to an apparent E2k pseudogene on chromosome 1p31. Northern analysis revealed the E2k gene to be ubiquitously expressed in peripheral tissues and brain. Interestingly, chromosome 14q24.3 has recently been reported to contain gene defects for an early-onset form of familial Alzheimer's disease and for Machado-Joseph disease. Future studies will be necessary to determine whether the E2K gene plays a role in either of these two disorders.

Development of seedless fruits mutants in citrus including tangerine (C. reticulata) and pummelo (C. grandis) through induced mutations and biotechnology

Energy Technology Data Exchange (ETDEWEB)

Somsri, S. [Horticulture Research Institute, Department of Agriculture (DOA), Chatuchak, Bangkok (Thailand); Jompook, P. [Department of Applied Radiation and Isotopes, Faculty of Science, Kasetsart University, Chatuchak, Bangkok (Thailand); Kanhom, P. [Phare Horticultural Research Center, Muang, Phare (Thailand); Thayamanont, P.; Meecharoen, S. [Pichit Horticultural Research Center, Muang, Pichit (Thailand); Kumcha, U. [Srisaket Horticultural Research Center, Muang, Srisaket (Thailand)

2009-05-15

The development of seedless fruit mutants in citrus, including Tangerine (C. reticulata) and Pummelo (C. grandis), through induced mutation and biotechnology was studied at the Gamma Irradiation Service and Nuclear Technology Center, Pichit and Phare Horticultural Research Center for 4 years (August 2000 to September 2004). The results showed successful induction of mutants with gamma irradiation using both chronic and acute procedures for pot plants, scions and in vitro plantlets of tangerine (Citrus reticulata var. Shogun and Sai Nam Puaeng) and pummelo (Citrus grandis viz. Kao Thong Dee). MS medium with 2 mgL{sup -1} of BA was found to be the most suitable medium for shoot proliferation. The seedlings were sub-cultured at least 4 times, and then they were treated with acute and chronic irradiation. Shoot induction from M{sub 1}V{sub 0} to M{sub 1}V{sub 4} generation was performed in basic MS medium with 2 mgL{sup -1} added BA. Rooting was induced in the M{sub 1}V{sub 4} in halfstrength MS enriched with BA 2 mgL{sup -1}. Later, the shoots were excised and grafted on mature plants or the plantlets directly transferred in the field and later the fruits from mature trees were evaluated for seedlessness in M{sub 1}V{sub 4} at Pichit and Phare Horticultural Research Center. (author)

Isolation of the human anionic glutathione S-transferase cDNA and the relation of its gene expression to estrogen-receptor content in primary breast cancer

International Nuclear Information System (INIS)

Moscow, J.A.; Townsend, A.J.; Goldsmith, M.E.; Whang-Peng, J.; Vickers, P.J.; Poisson, R.; Legault-Poisson, S.; Myers, C.E.; Cowan, K.H.

1988-01-01

The development of multidrug resistance in MCF7 human breast cancer cells is associated with overexpression of P-glycoprotein, changes in activities of several detoxication enzymes, and loss of hormone sensitivity and estrogen receptors (ERs). The authors have cloned the cDNA for one of the drug-detoxifying enzymes overexpressed in multidrug-resistant MCF7 cells (Adr R MCF7), the anionic isozyme of glutathione S-transferase (GSTπ). Hybridization with this GSTπ cDNA, GSTπ-1, demonstrated that increased GSTπ activity in Adr R MCF7 cells is associated with overexpression but not with amplification of the gene. They mapped the GSTπ gene to human chromosome 11q13 by in situ hybridization. Since multidrug resistance and GSTπ overexpression are associated with the loss of ERs in Adr R MCF7 cells, they examined several other breast cancer cell lines that were not selected for drug resistance. In each of these cell lines they found an inverse association between GSTπ expression and ER content. They also examined RNA from 21 primary breast cancers and found a similar association between GSTπ expression and ER content in vivo. The finding of similar patterns of expression of a drug-detoxifying enzyme and of ERs in vitro as well as in vivo suggests that ER-negative breast cancer cells may have greater protection against antineoplastic agents conferred by GSTπ than ER-positive tumors

Automation of cDNA Synthesis and Labelling Improves Reproducibility

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Daniel Klevebring

2009-01-01

Full Text Available Background. Several technologies, such as in-depth sequencing and microarrays, enable large-scale interrogation of genomes and transcriptomes. In this study, we asses reproducibility and throughput by moving all laboratory procedures to a robotic workstation, capable of handling superparamagnetic beads. Here, we describe a fully automated procedure for cDNA synthesis and labelling for microarrays, where the purification steps prior to and after labelling are based on precipitation of DNA on carboxylic acid-coated paramagnetic beads. Results. The fully automated procedure allows for samples arrayed on a microtiter plate to be processed in parallel without manual intervention and ensuring high reproducibility. We compare our results to a manual sample preparation procedure and, in addition, use a comprehensive reference dataset to show that the protocol described performs better than similar manual procedures. Conclusions. We demonstrate, in an automated gene expression microarray experiment, a reduced variance between replicates, resulting in an increase in the statistical power to detect differentially expressed genes, thus allowing smaller differences between samples to be identified. This protocol can with minor modifications be used to create cDNA libraries for other applications such as in-depth analysis using next-generation sequencing technologies.

Stimulatory effect of eucalyptus essential oil on the germination and early growth of Eucalyptus grandis seedlings Efeito estimulante do óleo essencial de eucalipto na germinação e crescimento inicial de mudas de Eucalyptus grandis

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Ricardo Bemfica Steffen

2010-12-01

Full Text Available

The use of essential oil and extracts from plants is becoming an efficient alternative in the biostimulation on growth and protection of plants. The aim of this work was to evaluate the use of leaf essential oil of Eucalyptus grandis on the germination and the  development of Eucalyptus grandis seedlings in nursery conditions. The eucalyptus seeds were exposed to the concentrations of 0, 10, 25, 50, 75 and 100 μL L-1 of the essential oil in controlled conditions. The eucalyptus seedlings were sprayed with 0, 20, 30, 40, 50 and 60 μL L-1 of the essential oil per plant, at intervals of seven days. The effect of this application on the seedling development were analyzed after 30 and 60 days. The results show that the germination was significantly higher when the seeds were exposed to 25 μL L-1 of the essential oil. The application of essential oil in the concentration of 30 and 40 μL L-1 provided higher growth of the aerial part and of the roots in greenhouse conditions, being an effective alternative to biostimulation the vegetative growth of eucalyptus seedlings.

doi: 10.4336/2010.pfb.30.63.199

Identification of eukaryotic open reading frames in metagenomic cDNA libraries made from environmental samples.

Science.gov (United States)

Grant, Susan; Grant, William D; Cowan, Don A; Jones, Brian E; Ma, Yanhe; Ventosa, Antonio; Heaphy, Shaun

2006-01-01

Here we describe the application of metagenomic technologies to construct cDNA libraries from RNA isolated from environmental samples. RNAlater (Ambion) was shown to stabilize RNA in environmental samples for periods of at least 3 months at -20 degrees C. Protocols for library construction were established on total RNA extracted from Acanthamoeba polyphaga trophozoites. The methodology was then used on algal mats from geothermal hot springs in Tengchong county, Yunnan Province, People's Republic of China, and activated sludge from a sewage treatment plant in Leicestershire, United Kingdom. The Tenchong libraries were dominated by RNA from prokaryotes, reflecting the mainly prokaryote microbial composition. The majority of these clones resulted from rRNA; only a few appeared to be derived from mRNA. In contrast, many clones from the activated sludge library had significant similarity to eukaryote mRNA-encoded protein sequences. A library was also made using polyadenylated RNA isolated from total RNA from activated sludge; many more clones in this library were related to eukaryotic mRNA sequences and proteins. Open reading frames (ORFs) up to 378 amino acids in size could be identified. Some resembled known proteins over their full length, e.g., 36% match to cystatin, 49% match to ribosomal protein L32, 63% match to ribosomal protein S16, 70% to CPC2 protein. The methodology described here permits the polyadenylated transcriptome to be isolated from environmental samples with no knowledge of the identity of the microorganisms in the sample or the necessity to culture them. It has many uses, including the identification of novel eukaryotic ORFs encoding proteins and enzymes.

Molecular cloning and characterization of an α-amylase cDNA highly expressed in major feeding stages of the coffee berry borer, Hypothenemus hampei.

Science.gov (United States)

Bezerra, C A; Macedo, L L P; Amorim, T M L; Santos, V O; Fragoso, R R; Lucena, W A; Meneguim, A M; Valencia-Jimenez, A; Engler, G; Silva, M C M; Albuquerque, E V S; Grossi-de-Sa, M F

2014-12-10

α-Amylases are common enzymes responsible for hydrolyzing starch. Insect-pests, whose larvae develop in seeds, rely obligatorily on α-amylase activity to digest starch, as their major food source. Considering the relevance of insect α-amylases and the natural α-amylase inhibitors present in seeds to protect from insect damage, we report here the molecular cloning and nucleotide sequence of the full-length AmyHha cDNA of the coffee berry borer, Hypothenemus hampei, a major insect-pest of coffee crops. The AmyHha sequence has 1879 bp, containing a 1458 bp open reading frame, which encodes a predicted protein with 485 amino acid residues, with a predicted molecular mass of 51.2 kDa. The deduced protein showed 55-79% identity to other insect α-amylases, including Anthonomus grandis, Ips typographus and Sitophilus oryzae α-amylases. In depth analysis revealed that the highly conserved three amino acid residues (Asp184, Glu220, and Asp285), which compose the catalytic site are also presented in AmyHha amylase. The AmyHha gene seems to be a single copy in the haploid genome and AmyHha transcription levels were found higher in L2 larvae and adult insects, both corresponding to major feeding phases. Modeling of the AmyHha predicted protein uncovered striking structural similarities to the Tenebrio molitor α-amylase also displaying the same amino acid residues involved in enzyme catalysis (Asp184, Glu220 and Asp285). Since AmyHha gene was mostly transcribed in the intestinal tract of H. hampei larvae, the cognate α-amylase could be considered a high valuable target to coffee bean insect control by biotechnological strategies. Copyright © 2014. Published by Elsevier B.V.

Coordinate regulation of stromelysin and collagenase genes determined with cDNA probes

International Nuclear Information System (INIS)

Frisch, S.M.; Clark, E.J.; Werb, Z.

1987-01-01

Secreted proteinases are required for tumor metastasis, angiogenesis, and tissue remodeling during wound healing and embryonic growth. Thus, the regulation of the genes of secreted proteinases may serve as an interesting model for growth-controlled genes in general. The authors studied the genes of the secreted proteinases stromelysin and collagenase by using molecularly cloned cDNAs from each proteinase. Stromelysin cDNA was cloned by differential screening of a total cDNA library from rabbit synovial cells treated with phorbol 12-myristate 13-acetate, which yielded a clone of 1.2 kilobase pairs; collagenase cDNA was obtained by cloning reverse transcripts of anti-collagenase-immunoadsorbed polysomal mRNA, which yielded a clone of 0.8 kilobase pairs. Stromelysin and collagenase mRNA species of 2.2 and 2.4 kilobases, respectively, were detected on hybridization blots of RNA from phorbol 12-myristate 13-acetate-treated but not untreated rabbit synovial cells. Expression of stromelysin mRNA was also induced in rabbit alveolar macrophages and rabbit brain capillary endothelial cells treated with phorbol 12-myristate 13-acetate. Stromelysin and collagenase mRNA were both induced by phorbol 12-myristate 13-acetate and cytochalasin B at a constant ratio of the two gene products; this suggest coordinate regulation. The fact that induction was blocked after inhibition of protein synthesis by cycloheximide implicates an indirect signal transduction pathway that requires new protein synthesis

Cloning of low dose radiation induced gene RIG1 by RACE based on non-cloned cDNA library

International Nuclear Information System (INIS)

Luo Ying; Sui Jianli; Tie Yi; Zhang Yuanping; Zhou Pingkun; Sun Zhixian

2001-01-01

Objective: To obtain full-length cDNA of radiation induced new gene RIG1 based on its EST fragment. Methods: Based on non-cloned cDNA library, enhanced nested RACE PCR and biotin-avidin labelled probe for magnetic bead purification was used to obtain full-length cDNA of RIG1. Results: About 1 kb of 3' end of RIG1 gene was successfully cloned by this set of methods and cloning of RIG1 5' end is proceeding well. Conclusion: The result is consistent with the design of experiment. This set of protocol is useful for cloning of full-length gene based on EST fragment

EXPRESSION PROFILING OF ESTROGENIC COMPOUNDS USING A SHEEPSHEAD MINNOW CDNA MACROARRAY

Science.gov (United States)

Larkin, Patrick, Leroy C. Folmar, Michael J. Hemmer, Arianna J. Poston and Nancy D. Denslow. 2003. Expression Profiling of Estrogenic Compounds Using a Sheepshead Minnow cDNA Macroarray. Environ. Health Perspect. 111(6):839-846. (ERL,GB 1171). A variety of anthropogenic c...

Cloning and expression of a cDNA coding for a human monocyte-derived plasminogen activator inhibitor.

Science.gov (United States)

Antalis, T M; Clark, M A; Barnes, T; Lehrbach, P R; Devine, P L; Schevzov, G; Goss, N H; Stephens, R W; Tolstoshev, P

1988-02-01

Human monocyte-derived plasminogen activator inhibitor (mPAI-2) was purified to homogeneity from the U937 cell line and partially sequenced. Oligonucleotide probes derived from this sequence were used to screen a cDNA library prepared from U937 cells. One positive clone was sequenced and contained most of the coding sequence as well as a long incomplete 3' untranslated region (1112 base pairs). This cDNA sequence was shown to encode mPAI-2 by hybrid-select translation. A cDNA clone encoding the remainder of the mPAI-2 mRNA was obtained by primer extension of U937 poly(A)+ RNA using a probe complementary to the mPAI-2 coding region. The coding sequence for mPAI-2 was placed under the control of the lambda PL promoter, and the protein expressed in Escherichia coli formed a complex with urokinase that could be detected immunologically. By nucleotide sequence analysis, mPAI-2 cDNA encodes a protein containing 415 amino acids with a predicted unglycosylated Mr of 46,543. The predicted amino acid sequence of mPAI-2 is very similar to placental PAI-2 (3 amino acid differences) and shows extensive homology with members of the serine protease inhibitor (serpin) superfamily. mPAI-2 was found to be more homologous to ovalbumin (37%) than the endothelial plasminogen activator inhibitor, PAI-1 (26%). Like ovalbumin, mPAI-2 appears to have no typical amino-terminal signal sequence. The 3' untranslated region of the mPAI-2 cDNA contains a putative regulatory sequence that has been associated with the inflammatory mediators.

Three human alcohol dehydrogenase subunits: cDNA structure and molecular and evolutionary divergence

International Nuclear Information System (INIS)

Ikuta, T.; Szeto, S.; Yoshida, A.

1986-01-01

Class I human alcohol dehydrogenase (ADH; alcohol:NAD + oxidoreductase, EC 1.1.1.1) consists of several homo- and heterodimers of α, β, and γ subunits that are governed by the ADH1, ADH2, and ADH3 loci. The authors previously cloned a full length of cDNA for the β subunit, and the complete sequence of 374 amino acid residues was established. cDNAs for the α and γ subunits were cloned and characterized. A human liver cDNA library, constructed in phage λgt11, was screened by using a synthetic oligonucleotide probe that was matched to the γ but not to the β sequence. Clone pUCADHγ21 and clone pUCADHα15L differed from β cDNA with respect to restriction sites and hybridization with the nucleotide probe. Clone pUCADHγ21 contained an insertion of 1.5 kilobase pairs (kbp) and encodes 374 amino acid residues compatible with the reported amino acid sequence of the γ subunit. Clone pUCADHα15L contained an insertion of 2.4 kbp and included nucleotide sequences that encode 374 amino acid residues for another subunit, the γ subunit. In addition, this clone contained the sequences that encode the COOH-terminal part of the β subunit at its extended 5' region. The amino acid sequences and coding regions of the cDNAs of the three subunits are very similar. A high degree of resemblance is observed also in their 3' noncoding regions. However, distinctive differences exist in the vicinity of the Zn-binding cysteine residue at position 46. Based on the cDNA sequences and the deduced amino acid sequences of the three subunits, their structural and evolutionary relationships are discussed

Potensi Kayu Perkakas dan Kayu Bakar Jenis Jati (Tectona grandis di Hutan Rakyat Desa Natah, Gunung Kidul

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Ris Hadi Purwanto

2009-07-01

Full Text Available Potentials  of Merchantable Timber and Firewod of Teak (Tectona grandis in Natah Village Community Forest , Gunungkidul The potential of merchantable timber and firewod of teak (Tectona grandis in Desa Natah community forest was estimated by developing allometric equations method. To establish the allometric equation 350 sample trees were measured to determine the relationships between tree height (H and diameter breast height (D. Thirty trees of various sizes were cut to measure the merchantable timber and firewood volume. The raw merchantable timber volume of teak in the community forest was defined as the ligneous material contained in the bole and branches which both with a diameter of at least 10cm. The result showed that D (taken at about 1.3m above the ground was a good predictor of H with r2 over 0.9672. When D was combined with H, r2 was improved somewhat for the merchantable timber volume, suggesting the growth patterns of tree dimensions were colesy interdependent. A standing stock of the merchantable timber and firewood volume of teak in the community forest was then estimated based on the allometric relations. Proportions of the merchantable timber and firewood volume were 66.91% and 33.09% of total wood volume per tree, respectively. The potential of merchantable timber and firewood volume in these community forest were 13.501 m3/ha and 8.686 m3/ha, respectively, with a basal area of 1.887 m2/ha. Based on the basal area, Desa Natah community forests of teak could be classified into extremely sparse of stands category.

EFEITO DA TERMORRETIFICAÇÃO NAS PROPRIEDADES MECÂNICAS DAS MADEIRAS DE Pinus taeda E Eucalyptus grandis

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Karina Soares Modes

2017-01-01

Full Text Available This study investigated the effect of thermal treatment, employed under two treatment conditions on mechanical properties of wood of Pinus taeda L. and Eucalyptus grandis W. Hill ex Maiden. From each species, three 25-year tress were sampled, and from each one, it was taken the first 2-meter long log, which was obtained from two boards of size 7,0 x 20,0 cm (thickness x width respectively diametrically opposite from where 30 specimens were saw for each treatment. At the first one, called the combined one, the woods were subjected to heat treatment by autoclaving at 130 / ± 3°C and pressure of 2 kgf / cm ² for 3 hours and, after a conditioning period, subjected to heat in an electric oven at 160 / ± 1°C for the same period. The second treatment consisted only of heat treatment in oven. It was also evaluated the pieces of wood without treatment (control. The mechanical properties were evaluated by means of tests for determining the modulus of elasticity and rupture in bending, maximum resistance to compression parallel to the grain and Janka hardness according to ASTM D 143 (1995, and the impact resistance according to ABNT NBR 7190 (1997. For Pinus taeda wood, it was observed that treatment in an oven gave the worst outcomes, both due to the reduction in the values of supported load of a greater number of mechanical properties evaluated, but also as compared to the lowest increments in resistance when it was observed an increase to the same ones with heat treatment. In Eucalyptus grandis, the combined treatment decreased the greatest number of mechanical properties of wood.

Pontoscolex corethrurus (Annelida: Oligochaeta indicador de la calidad del suelo en sitios de Eucalyptus grandis (Myrtacea con manejo tumba y quema Pontoscolex corethrurus (Annelidae: Oligochaeta soil quality indicator in Eucalyptus grandis (Myrtacea sites with slash and burn management

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Sheila Uribe

2012-12-01

Full Text Available La presencia de oligoquetos en los ecosistemas puede indicar fertilidad del suelo, ya que estos organismos transportan, mezclan y entierran los residuos vegetales de la superficie al interior del suelo. Se caracterizó la comunidad de oligoquetos bajo sitios con diferentes periodos de establecimiento y manejo de plantaciones de Eucalyptus grandis, sin vegetación (SV, con cinco años en producción (Euc y vegetación secundaria con 15 años (Acah que han pasado por el proceso de tumba y quema en suelos de Acrisol en Huimanguillo, Tabasco; y se analizaron las propiedades físico-químicas del suelo (D.A., humedad, textura, pH, Ntot, MO, P, K, CIC. La recolecta de lombrices se realizó al finalizar las lluvias (agosto-octubre 2007. Se muestreó en tres parcelas con seis réplicas en cada una. Se encontró que los suelos tenían pH de 3.0-4.5 en los primeros 30cm de profundidad. Los contenidos de materia orgánica (MO y nitrógeno total (Ntot fueron significativamente menores en los sitios SV (6-8% y 0.19-0.22% respectivamente que en Euc y Acah (MO=9-11%; el Ntot=0.27-0.33%. La especie Pontoscolex corethrurus domino en toda el área, presentando mayores densidades y biomasas en Euc (164.4ind/m² y 36.8g/m² respectivamente y Acah (138.7ind/m² y 19.1g/m² respectivamente, mientras que en SV sus poblaciones fueron reducidas en un 80%. Se encontró que el sistema Acah sigue presentando rasgos de un sistema perturbado, al no recuperar fácilmente la diversidad de oligoquetos y las concentraciones de nutrientes disponibles en el sueloSoil burning has been used in agricultural and forestry systems as a fundamental technique to clean the land and add some nutrients to the soil. In addition, earthworms are known to promote various soil functions since they contribute to aeration and organic matter and nutrients availability to other soil organisms. This study evaluated the effects of tropical forest crops management with presence-absence of Eucalyptus

Optimal management and productivity of Eucalyptus grandis on former phosphate mined and citrus lands in central and southern Florida: influence of genetics and spacing

Science.gov (United States)

Kyle W. Fabbro; Donald L. Rockwood

2016-01-01

Eucalyptus short rotation woody crops (SRWC) with superior genotypes are promising in central and south Florida due to their fast growth, freeze resilience, coppicing ability, and site tolerance. Four Eucalyptus grandis cultivars, E.nergy™ G1, G2, G3, and/or G5, were established in 2009 at varying planting densities on a...

Modification of the original color of the Eucalyptus grandis wood by heat treatments

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Rosilei Aparecida Garcia

2014-09-01

Full Text Available The objective of this study was to determine the modification of original color of Eucalyptus grandis Hill ex. Maiden wood after heat-treatment. Wood samples were heat-treated under different temperatures (180, 200, 215 and 230ºC and time conditions (15 minutes, 2 and 4 hours. Color analysis were performed on the CIE L*a*b* system by using a Color Eye XTH-X-Rite 200d spectrophotometer. All heat treatments promoted an alteration of the original color of wood. Heat-treated woods presented lower L* (lightness values than untreated wood (control, characterizing the wood darkness, mainly for more severe conditions of temperature and time. Chromatic coordinates (a* and b* showed different behaviors depending on the temperature-time combination. The modification of the original color of the wood allowed the creation of new color patterns, which can add greater value to the studied wood.

Eradication of the cotton boll weevil (Anthonomus grandis) in the United States - A successful multi-regional approach

International Nuclear Information System (INIS)

Cunningham, Gary L.; Grefenstette, William J.

2000-01-01

The cotton boll weevil, Anthonomus grandis Boheman, is believed to have entered the US from Mexico and was first detected in South Texas in 1892. Since that time, the pest has spread throughout most of the nation's cotton-producing areas and has become the industry's number one nemesis. More than US$13 billion in economic losses have occurred since its introduction, with recent annual expenditures of more than US$300 million for control costs alone. Although the weevil has been eradicated from over 4 million acres, its presence in non-programme areas continues to dictate production practices within the mid-south, Texas and Oklahoma

cDNA, deduced polypeptide structure and chromosomal assignment of human pulmonary surfactant proteolipid, SPL(pVal)

International Nuclear Information System (INIS)

Glasser, S.W.; Korfhagen, T.R.; Weaver, T.E.; Clark, J.C.; Pilot-Matias, T.; Meuth, J.; Fox, J.L.; Whitsett, J.A.

1988-01-01

In hyaline membrane disease of premature infants, lack of surfactant leads to pulmonary atelectasis and respiratory distress. Hydrophobic surfactant proteins of M/sub r/ = 5000-14,000 have been isolated from mammalian surfactants which enhance the rate of spreading and the surface tension lowering properties of phospholipids during dynamic compression. The authors have characterized the amino-terminal amino acid sequence of pulmonary proteolipids from ether/ethanol extracts of bovine, canine, and human surfactant. Two distinct peptides were identified and termed SPL(pVal) and SPL(Phe). An oligonucleotide probe based on the valine-rich amino-terminal amino acid sequence of SPL(pVal) was utilized to isolate cDNA and genomic DNA encoding the human protein, termed surfactant proteolipid SPL(pVal) on the basis of its unique polyvaline domain. The primary structure of a precursor protein of 20,870 daltons, containing the SPL(pVal) peptide, was deduced from the nucleotide sequence of the cDNAs. Hybrid-arrested translation and immunoprecipitation of labeled translation products of human mRNA demonstrated a precursor protein, the active hydrophobic peptide being produced by proteolytic processing. Two classes of cDNAs encoding SPL(pVal) were identified. Human SPL(pVal) mRNA was more abundant in the adult than in fetal lung. The SPL(pVal) gene locus was assigned to chromosome 8

cDNA, genomic sequence cloning and analysis of the ribosomal ...

African Journals Online (AJOL)

Ribosomal protein L37A (RPL37A) is a component of 60S large ribosomal subunit encoded by the RPL37A gene, which belongs to the family of ribosomal L37AE proteins, located in the cytoplasm. The complementary deoxyribonucleic acid (cDNA) and the genomic sequence of RPL37A were cloned successfully from giant ...

Observation of intermittency in gene expression on cDNA microarrays

CERN Document Server

Peterson, L E

2002-01-01

We used scaled factorial moments to search for intermittency in the log expression ratios (LERs) for thousands of genes spotted on cDNA microarrays (gene chips). Results indicate varying levels of intermittency in gene expression. The observation of intermittency in the data analyzed provides a complimentary handle on moderately expressed genes, generally not tackled by conventional techniques.

cDNA sequence and tissue expression analysis of glucokinase from ...

African Journals Online (AJOL)

Yomi

2012-01-10

Jan 10, 2012 ... distribution of GK mRNA in brain, mesenteric adipose tissue, spleen, white muscle and liver of grass ... expression profile of GK mRNA in liver normalized with β-actin level was 31, 454 and 649-fold compared .... Primers and expected products used for GK gene cDNA RT-PCR, RACE and real-time PCR.

Crecimiento y propiedades fisico-mecanicas de la madera de teca (Tectona grandis de 17 anos de edad en San Joaquin de Abangares, Costa Rica

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Fernando Castro

2000-01-01

Full Text Available Growth and physical-mechanical properties of 17 years old teak (Tectona grandis growing in San Joaquin, Abangares, Costa Rica. The national and international market of forest products, from certifieddad. managment forests and plantations, is increasingly more demanding as to the standards and characteristics of high quality. The physicalmecanicas mechanical properties of the teak (Tectona grandis, growing in San Joaqufn de Abangares, Costa Rica, at 30 m and 100 16´ north latitude, are determined according to the ASTM standard Destudiadas 143-83. The physical properties of teak timber studied were: basic specific weight, radial, tangential and volumetric contractions, contraction ratio, dry and green density and point of fiber saturation. The mechanical properties studied were: static flexure, shear, hardness, parallel and perpendicular compression. Also included were comparisons with teak timber harvested in other places and latitudes, as well as other hardwood species. The coefficient of variation of the basic specific weight of the San Joaquin de Abangares teak (3.4% is half that of the Quepos teak (7% and almost one third of the average of 50 species (10%, which is an...

Characterization of cDNA encoding human placental anticoagulant protein (PP4): Homology with the lipocortin family

International Nuclear Information System (INIS)

Grundmann, U.; Abel, K.J.; Bohn, H.; Loebermann, H.; Lottspeich, F.; Kuepper, H.

1988-01-01

A cDNA library prepared from human placenta was screened for sequences encoding the placental protein 4 (PP4). PP4 is an anticoagulant protein that acts as an indirect inhibitor of the thromboplastin-specific complex, which is involved in the blood coagulation cascade. Partial amino acid sequence information from PP4-derived cyanogen bromide fragments was used to design three oligonucleotide probes for screening the library. From 10 6 independent recombinants, 18 clones were identified that hybridized to all three probes. These 18 recombinants contained cDNA inserts encoding a protein of 320 amino acid residues. In addition to the PP4 cDNA the authors identified 9 other recombinants encoding a protein with considerable similarity (74%) to PP4, which was termed PP4-X. PP4 and PP4-X belong to the lipocortin family, as judged by their homology to lipocortin I and calpactin I

Isolation and mapping of a C3'H gene (CYP98A49) from globe artichoke, and its expression upon UV-C stress

NARCIS (Netherlands)

Moglia, A.; Comino, C.; Portis, E.; Acquadro, A.; Vos, de C.H.; Beekwilder, M.J.; Lanteri, S.

2009-01-01

Globe artichoke represents a natural source of phenolic compounds with dicaffeoylquinic acids along with their biosynthetic precursor chlorogenic acid (5-caffeoylquinic acid) as the predominant molecules. We report the isolation and characterization of a full-length cDNA and promoter of a globe

Preparation of Proper Immunogen by Cloning and Stable Expression of cDNA coding for Human Hematopoietic Stem Cell Marker CD34 in NIH-3T3 Mouse Fibroblast Cell Line

Science.gov (United States)

Shafaghat, Farzaneh; Abbasi-Kenarsari, Hajar; Majidi, Jafar; Movassaghpour, Ali Akbar; Shanehbandi, Dariush; Kazemi, Tohid

2015-01-01

Purpose: Transmembrane CD34 glycoprotein is the most important marker for identification, isolation and enumeration of hematopoietic stem cells (HSCs). We aimed in this study to clone the cDNA coding for human CD34 from KG1a cell line and stably express in mouse fibroblast cell line NIH-3T3. Such artificial cell line could be useful as proper immunogen for production of mouse monoclonal antibodies. Methods: CD34 cDNA was cloned from KG1a cell line after total RNA extraction and cDNA synthesis. Pfu DNA polymerase-amplified specific band was ligated to pGEMT-easy TA-cloning vector and sub-cloned in pCMV6-Neo expression vector. After transfection of NIH-3T3 cells using 3 μg of recombinant construct and 6 μl of JetPEI transfection reagent, stable expression was obtained by selection of cells by G418 antibiotic and confirmed by surface flow cytometry. Results: 1158 bp specific band was aligned completely to reference sequence in NCBI database corresponding to long isoform of human CD34. Transient and stable expression of human CD34 on transfected NIH-3T3 mouse fibroblast cells was achieved (25% and 95%, respectively) as shown by flow cytometry. Conclusion: Cloning and stable expression of human CD34 cDNA was successfully performed and validated by standard flow cytometric analysis. Due to murine origin of NIH-3T3 cell line, CD34-expressing NIH-3T3 cells could be useful as immunogen in production of diagnostic monoclonal antibodies against human CD34. This approach could bypass the need for purification of recombinant proteins produced in eukaryotic expression systems. PMID:25789221

Cdna cloning and expression analyses of the isoflavone reductase-like gene of dendrobium officinale

International Nuclear Information System (INIS)

Qian, X.; Xu, S.Z.

2015-01-01

The full length of the isoflavone reductase-like gene (IRL) cDNA of Dendrobium officinale was cloned by using reverse transcription (RT) PCR combined with cDNA library, the IRL function was identified by Bioinformatics and prokaryotic expression analyses, and the IRL expression levels in the organs and tissues of D. officinale plants with different ages were determined by using real-time quantitative PCR (RT-qPCR). The results indicated that the full length of the cDNA of D. officinale IRL, DoIRL, was 1238 bp (accession no. KJ661023). Its open reading frame (ORF) was 930 bp which encoded 309 amino acids with a predicted molecular mass of 34 kDa, the 5 untranslated region (UTR) was 61 bp and the 3 UTR containing a poly (A) tail was 247 bp. The deduced amino acid sequence of DoIRL, DoIRL, was forecast to contain a NAD(P)H-binding motif (GGTGYIG) in the N-terminal region, two conserved N-glycosylation sites, a conserved nitrogen metabolite repression regulator (NmrA) domain and a phenylcoumaran benzylic ether reductase (PCBER) domain, to hold the nearest phylogenetic relationship with the PCBER of Striga asiatica, and to share both 73% identity with the isoflavone reductases-like (IRLs) of Cucumis sativus and Striga asiatica. In Escherichia coli 'BL21' cells, the DoIRL cDNA expression produced a protein band holding the predicted molecular mass of 34 kDa. DoIRL expressed in all organs and tissues of D. officinale plants with different ages at comparatively low levels, and the expression level in the leaves of the two-year-old plants was the highest. (author)