WorldWideScience
 
 
1

Use of synthetic oligonucleotide DNA probes for identification and direct detection of Bacteroides forsythus in plaque samples.  

UK PubMed Central (United Kingdom)

Oligonucleotide DNA probes complementary to the hypervariable region of the 16S rRNA of Bacteroides forsythus were tested for their specificity and sensitivity against reference and clinical isolates...Full Text Available

1991-10-01

3

Nucleotide sequence of a Euglena gracilis chloroplast gene coding for the 16S rRNA: homologies to E. coli and Zea mays chloroplast 16S rRNA.  

UK PubMed Central (United Kingdom)

The nucleotide sequence of 16S rDNA from Euglena gracilis chloroplasts has been determined representing the first complete sequence of an algal chloroplast rRNA gene. The structural part of the 16S...Full Text Available

1982-10-25

4

chipD: a web tool to design oligonucleotide probes for high-density tiling arrays  

UK PubMed Central (United Kingdom)

chipD is a web server that facilitates design of DNA oligonucleotide probes for high-density tiling arrays, which can be used in a number of genomic applications such as ChIP-chip or gene-expression...Full Text Available

2010-07-01

5

Simultaneous rapid chemical synthesis of over one hundred oligonucleotides on a microscale  

UK PubMed Central (United Kingdom)

An inexpensive, extremely rapid manual method for simultaneous synthesis of large numbers of oligodeoxyribonucleotides on 50 or 150 nanomole scale is described. The oligonucleotides are assembled in...Full Text Available

1984-04-01

6

Biotin-labeled hairpin oligonucleotides: probes to detect double-strand breaks in DNA in apoptotic cells.  

UK PubMed Central (United Kingdom)

Hairpin oligonucleotides were synthesized with stems ending in a double-stranded structure, which can be ligated to double-strand breaks in DNA, and with loops that contain nucleotides modified by the...Full Text Available

1998-04-01

8

Emergence of Klebsiella pneumoniae Coproducing KPC-2 and 16S rRNA Methylase ArmA in Poland ?  

UK PubMed Central (United Kingdom)

A Klebsiella pneumoniae epidemic strain that coproduced carbapenemase KPC-2 (K. pneumoniae carbapenemase 2) and 16S rRNA methylase ArmA has emerged in Poland. Four...Full Text Available

2011-01-01

9

Hybridization with synthetic oligonucleotides  

Science.gov (United States)

Procedures are described for the use of synthetic oligonucleotides for Southern blot experiments and gene bank screening, and the effect of various mismatches on the efficiency of hybridization is demonstrated. The following topics are discussed: sensitivity vs. specificity, hybridization of a 12-mer to the lambda endolysin gene; hybridization of oligonucleotide probes to the E. coli lac operator; hybridization of synthetic probes to the CYC1 gene of yeast; and cloning eucaryotic genes. (HLW)

1978-01-01

10

211At-Rh(16-S4-diol) as a starting complex for preparing an astatine-labeled radiopharmaceutical  

British Library Electronic Table of Contents (United Kingdom)

A new procedure for preparing an 211At-labeled radiopharmaceutical is suggested. The 211At? anion forms a strong bond with the Rh3+ cation incorporated in the complex with a thiother ligand, 1,5,9,13-tetrathiacyclohexadecane-3,11-diol (16-S4-diol). The reaction conditions are optimized with 131I as astatine analog. The complexes are studied by paper electrophoresis, ion exchange, and thin-layer chromatography. The kinetics of the addition of the 131I? anion to Rh(16-S4-diol) and the dependence of the yield of the forming complex 131I-Rh(16-S4-diol) on the temperature, solution acidity, and reactant concentrations are examined. Taking into account the results obtained, the complex 211At-Rh(16-S4-diol) is prepared by adding astatide (211 At?) to equivalent amounts of RhCl3 and the tetrathioe...

2008-01-01

11

"2"1"1At-Rh(16-S4-diol) - initial complex of radiopharmaceuticals labelled with astatine  

International Nuclear Information System (INIS)

Paper describes a new procedure to synthesize a pharmaceutical containing "2"1"1At. "2"1"1At"- anion was found to bind strongly with Rh"3"+ cation complexing with a thio-ether ligand - 1,5,9,13-tetratiacyclohexadecane-3,11-diol (16-S4-diol). One made use of "1"3"1I isotope to seek for the optimal conditions of the reaction. The derived complexes were studied by means of the paper electrophoresis, the ion exchange, the surface chromatography. One studied the kinetics of "1"3"1I"- anion binding with Rh(16-S4-diol), the dependence of the yield of "1"3"1I-Rh(16-S4-diol) forming complex on the temperature. the acidity of the reaction mixture and the concentration of the reagents. On the basis of the determined conditions of the synthesis one prepared "2"1"1At-Rh(16-S4-diol) complex by more astatide ("2"1"1At"-) to the ...

13

Carbohydrates/nucleosides/RNA-DNA-ligand interactions  

Energy Technology Data Exchange (ETDEWEB)

Carbohydrate and nucleotide structural determination using modern spectroscopic techniques is dependent on our ability to label oligonucleotides and oligosaccharides with stable isotopes. Uniform Carbon 13 and Nitrogen 15 labeling of oligonucleotides is important to present-day efforts, which are focused on determining the structure of relatively small oligosaccharides and oligonucleotides, which form the elements of larger structures. Because of the relatively recent interest in three-dimensional structure, the development of techniques used to label them has lagged behind parallel techniques used to label peptides and proteins. Therefore, this group`s discussion focused primarily on problems faced today in obtaining oligonucleotides labeled uniformly with carbon 13 and nitrogen 15.

1994-12-01

14

Antimicrobial activity and mechanism of action of Nu-3, a protonated modified nucleotide  

UK PubMed Central (United Kingdom)

Background"Nubiotics" are synthetic oligonucleotides and nucleotides with nuclease-resistant backbones, and are fully protonated for enhanced ability to be taken up by bacterial...Full Text Available

16

Large T1 oligonucleotides of Moloney leukemia virus missing in an env gene recombinant, HIX, are present on an intracellular 21S Moloney viral RNA species.  

UK PubMed Central (United Kingdom)

HIX, a recombinant derived from Moloney leukemia virus, has an envelope glycoprotein different from that of the Moloney virus. HIX and Moloney viruses share the majority of the large T1 oligonucleotides...Full Text Available

1978-06-01

17

Isolation and sequence determination of 5'-terminal oligonucleotide fragments of RNA transcripts synthesized by bacteriophage T3-induced RNA polymerase from T3 DNA.  

UK PubMed Central (United Kingdom)

The nucleotide sequence of the 5'-terminal oligonucleotides produced by pancreatic RNase digestion of bacteriophage T3 RNA polymerase (EC 2.7.7.6) transcripts of T3 DNA has been determined. The sequence...Full Text Available

1980-07-01

18

Discrimination of Listeria monocytogenes from other Listeria species by ligase chain reaction.  

UK PubMed Central (United Kingdom)

A ligase chain reaction assay based on a single-base-pair difference in the V9 region of the 16S rRNA gene (16S rDNA) was developed to distinguish between Listeria monocytogenes and other Listeria species....Full Text Available

1992-11-01

19

Use of an alkaline phosphatase-labeled synthetic oligonucleotide probe for detection of Campylobacter jejuni and Campylobacter coli.  

UK PubMed Central (United Kingdom)

A commercially available synthetic nucleic acid probe (SNAP) conjugated to alkaline phosphatase was compared with standard culture techniques for detecting Campylobacter species. The SNAP was able to...Full Text Available

1990-07-01

20

Prevention of guanine modification and chain cleavage during the solid phase synthesis of oligonucleotides using phosphoramidite derivatives.  

UK PubMed Central (United Kingdom)

Phosphoramidite reagents can phosphitylate guanine bases at the O6-position during solid phase synthesis and serious chain cleavage occurs if the base phosphitylation is not eliminated before the iodine/water...Full Text Available

1986-08-26

 
 
 
 
21

Modification of guanine bases by nucleoside phosphoramidite reagents during the solid phase synthesis of oligonucleotides.  

UK PubMed Central (United Kingdom)

Nucleoside 3'-phosphoramidite and chlorophosphite reagents have been found to react with the lactam function of guanine. This reaction caused unsatisfactory results when oligodeoxyribonucleotides containing...Full Text Available

1985-09-25

22

Intracellular delivery of an antisense?oligonucleotide via endocytosis of a G protein-coupled receptor  

UK PubMed Central (United Kingdom)

Gastrin-releasing peptide receptor (GRPR), a member of the G protein-coupled receptor superfamily, has been utilized for receptor-mediated targeting of imaging and therapeutic agents; here we extend...Full Text Available

2010-10-01

23

High-density multiplex detection of nucleic acid sequences: oligonucleotide ligation assay and sequence-coded separation.  

UK PubMed Central (United Kingdom)

We describe a non-isotopic, semi-automated method for large-scale multiplex analysis of nucleic acid sequences, using the cystic fibrosis transmembrane regulator (CFTR) gene as an example. Products...Full Text Available

1994-10-25

24

5-Fluorodeoxyuridine as an alternative to the synthesis of mixed hybridization probes for the detection of specific gene sequences.  

UK PubMed Central (United Kingdom)

Synthetic complementary oligonucleotides are useful hybridization probes for the detection of mRNAs and genes encoding proteins for which only a partial amino acid sequence is known. Usually this involves...Full Text Available

1988-03-01

25

Worlds within worlds: evolution of the vertebrate gut microbiota  

UK PubMed Central (United Kingdom)

Here we use published 16S rRNA gene sequences to compare the bacterial assemblages associated with humans, other mammals, other metazoa, and free-living microbial communities spanning a range...Full Text Available

2008-10-01

26

PCR-based method for targeting 16S-23S rRNA intergenic spacer regions among Vibrio species  

UK PubMed Central (United Kingdom)

BackgroundThe genus Vibrio is a diverse group of Gram-negative bacteria comprised of 74 species. Furthermore, the genus has and is expected to continue expanding...Full Text Available

27

Influence of Species Specificity and Other Factors on Bacteria Associated with the Coral Stylophora pistillata in Taiwan ?  

UK PubMed Central (United Kingdom)

Species of bacteria associated with Stylophora pistillata were determined by analyses of 16S ribosomal genes. Coral samples were taken from two distinct sites at Kenting, in the far...Full Text Available

2009-12-01

28

Improved Understanding of the Bacterial Vaginal Microbiota of Women before and after Probiotic Instillation  

UK PubMed Central (United Kingdom)

The vaginal bacterial microbiota of 19 premenopausal women was examined by PCR-denaturing gradient gel electrophoresis (DGGE) and sequencing of the V2-V3 region of the 16S rRNA gene. Ten of the women...Full Text Available

2003-01-01

29

Detection of Atopobium vaginae in Postmenopausal Women by Cultivation-Independent Methods Warrants Further Investigation  

UK PubMed Central (United Kingdom)

We sequenced 16S rRNA genes from the vaginal swab contents of a postmenopausal woman with asymptomatic bacterial vaginosis (BV). Sequences from Atopobium vaginae were the most commonly...Full Text Available

2004-04-01

30

Association of Unique, Isolated Treponemes with Bovine Digital Dermatitis Lesions?  

UK PubMed Central (United Kingdom)

This study used a PCR-based approach targeting 16S rRNA gene fragments to determine the occurrence and association of the three bovine digital dermatitis (BDD) treponeme phylogroups within lesions found...Full Text Available

2009-03-01

31

"2"1"1At-Rh(16-S4-diol) complex as a precursor for astatine radiopharmaceuticals  

International Nuclear Information System (INIS)

"2"1"1At is one of the most promising radionuclides in #alpha#-radioimmunotherapy (#alpha#-RIT). Unfortunately, biomolecules labeled by direct electrophilic astatination are unstable due to the rapid loss of "2"1"1At under both in vitro and in vivo conditions. The present paper describes the results of our studies on attaching At"- to the rhodium(III) complex with thioether ligand: 1,5,9,13-etrathiacyclohexadecane-3,11-diol (16-S4-diol). Rh"3"+ was chosen as a moderately soft metal cation which should form very strong bonds with soft At"- anions, but first of all because of the kinetic inertness of low spin rhodium(III) d"6 complexes. The 16-S4-diol ligand was selected due to formation of stable complexes with Rh"3"+. The experiments related to optimization of the reaction conditions were performed with the "1"3"1I, basing on a chemical similarity of I"- to At"-. The experiments with "2"1"1At were then ...

32

Electrochemical detection of DNA hybridization based on silver-enhanced gold nanoparticle label  

Energy Technology Data Exchange (ETDEWEB)

An electrochemical detection method for analyzing sequence-specific DNA using gold nanoparticle DNA probes and subsequent signal amplification step by silver enhancement is described. The assay relies on the electrostatic adsorption of target oligonucleotides onto the sensing surface of the glassy carbon electrode (GCE) and its hybridization to the gold nanoparticle-labeled oligonucleotides DNA probe. After silver deposition onto gold nanoparticles, binding events between probe and target were monitored by the differential pulse voltammetry (DPV) signal of the large number of silver atoms anchored on the hybrids at the electrode surface. The signal intensity difference permits to distinguish between the match of two perfectly matched DNA strands and the near-perfect match where just one base pair was wrong. Coupled with this 'nanoparticle-promoted' reduction of silver signal amplification method, the sensitivity of this ...

2002-10-03

33

Mutations at the cysteine codons of the recA gene of Escherichia coli  

International Nuclear Information System (INIS)

Each of the three cysteine residues in the Escherichia coli RecA protein was replaced with a number of other amino acids. To do this, each cysteine codon was first converted to a chain-terminating amber codon by oligonucleotide-directed mutagenesis. These amber mutants were then either assayed for function in different suppressor strains or reverted by a second round of mutagenesis with oligonucleotides that had random sequences at the amber codon. Thirty-three different amino acid substitutions were obtained. Mutants were tested for three functions of RecA: survival following UV irradiation, homologous recombination, and induction of the SOS response. It was found that although none of the cysteines is essential for activity, mutations at each of these positions can affect one or more of the activities of RecA, depending on the particular amino acid substitution. In addition, the cysteine at position 116 appears to be involved in the ...

34

Oxidation of Elemental Sulfur by Fusarium solani Strain THIF01 Harboring Endobacterium Bradyrhizobium sp.  

British Library Electronic Table of Contents (United Kingdom)

Nineteen fungal strains having an ability to oxidize elemental sulfur in mineral salts medium were isolated from deteriorated sandstones of Angkor monuments. These fungi formed clearing zone on agar medium supplemented with powder sulfur due to the dissolution of sulfur. Representative of the isolates, strain THIF01, was identified as Fusarium solani on the basis of morphological characteristics and phylogenetic analyses. PCR amplification targeting 16S rRNA gene and analyses of full 16S rRNA gene sequence indicated strain THIF01 harbors an endobacterium Bradyrhizobium sp.; however, involvement of the bacterium in the sulfur oxidation is still unclear. Strain THIF01 oxidized elemental sulfur to thiosulfate and then sulfate. Germination of the spores of strain THIF01 was observed in a liqui...

2010-01-01

35

Distinctive Responses of Metabolically Active Microbiota to Acidification in a Thermophilic Anaerobic Digester  

British Library Electronic Table of Contents (United Kingdom)

Acidification is one of the most common and serious problems inducing process failure in anaerobic digesters. The production of volatile fatty acids (VFAs) mainly triggers acidic shock. However, little is known about the bacteria involved in the processes of acidogenic metabolism, such as fermentation and reductive acetogenesis. Here, the metabolic responses of a methanogenic community to the acidification and resulting process deterioration were investigated using transcriptional profiling of both the 16S rRNA and formyltetrahydrofolate synthetase (FTHFS) genes. The 16S rRNA-based analyses demonstrated that the dynamic shift of bacterial populations was closely correlated with reactor performance, especially with VFA accumulation levels. The pH drop accompanied by an increase in VFAs stim...

2011-01-01

36

Termination of transcription by bacteriophage T3 RNA polymerase: homogeneous 3'-terminal oligonucleotide sequence of in vitro T3 RNA polymerase transcripts.  

UK PubMed Central (United Kingdom)

RNA was synthesized in vitro from a T3 DNA template by T3 RNA polymerase and subsequently separated into seven discrete size classes (molecular weights ranging between 0.21 x 10(6) and 6.2 x 10(6))...Full Text Available

1979-10-01

37

Sustainable phosphorous fertilisation of potatoes (Potato CHIPS)  

Environmental Research Database

DescriptionThis project has two independent aims: (1) to investigate the use of struvite as an alternative to chemical P fertilisers and (2) to develop an oligonucleotide microarray to monitor the P status of the potato crop. The UK horticultural and agricultural industries rely on large inputs of phosphate (P) fertilisers to maintain crop yields and quality. However, the use of non-renewable, chemical P fertilisers is unsustainable, and the alternatives to chemical P-fertilisers must be identified as an [continued...

2008-01-31

38

Isolation, sequence, and expression in Escherichia coli of an unusual thioredoxin gene from the cyanobacterium Anabaena sp. strain PCC 7120.  

UK PubMed Central (United Kingdom)

Two sequences with homology to a thioredoxin oligonucleotide probe were detected by Southern blot analysis of Anabaena sp. strain PCC 7120 genomic DNA. One of the sequences was shown to code for a protein...Full Text Available

1989-01-01

39

Insights into finding a mismatch through the structure of a mispaired DNA bound by a rhodium intercalator  

UK PubMed Central (United Kingdom)

We report the 1.1-Å resolution crystal structure of a bulky rhodium complex bound to two different DNA sites, mismatched and matched in the oligonucleotide 5′-(dCGGAAATTCCCG)2-3′....Full Text Available

2007-01-09

40

Microbial dynamics in upflow anaerobic sludge blanket (UASB) bioreactor granules in response to short-term changes in substrate feed  

Science.gov (United States)

The complexity and diversity of the microbial communities in biogranules from an upflow anaerobic sludge blanket (UASB) bioreactor were determined in response to short-term changes in substrate feeds. The reactor was fed simulated brewery wastewater (SBWW) (70% ethanol, 15% acetate, 15% propionate) for 1.5 months (phase 1), acetate / sulfate for 2 months (phase 2), acetate-alone for 3 months (phase 3), and then a return to SBWW for 2 months (phase 4). Performance of the reactor remained relatively stable throughout the experiment as shown by COD removal and gas production. 16S rDNA, methanogen-associated mcrA and sulfate reducer-associated dsrAB genes were PCR amplified, then cloned and sequenced. Sequence analysis of 16S clone libraries showed a relatively simple community composed mainly of the methanogenic Archaea (Methanobacterium and Methanosaeta), members of the Green Non-Sulfur (Chloroflexi) ...

2010-08-01

 
 
 
 
41

Detection of Atopobium vaginae in postmenopausal women by cultivation-independent methods warrants further investigation.  

Science.gov (United States)

We sequenced 16S rRNA genes from the vaginal swab contents of a postmenopausal woman with asymptomatic bacterial vaginosis (BV). Sequences from Atopobium vaginae were the most commonly detected. In a survey of 35 other postmenopausal women, this organism was detected in 44% with BV but not in any subjects deemed healthy. PMID:15071062

2004-04-01

42

User-friendly algorithms for estimating completeness and diversity in randomized protein-encoding libraries.  

Science.gov (United States)

Directed evolution of proteins depends on the production of molecular diversity by random mutagenesis. While a number of methods have been developed for introducing this diversity, the best ways to sample it are not always clear. Here we used simple statistics to analyse completeness and diversity in randomized libraries generated by oligonucleotide-directed mutagenesis, error-prone polymerase chain reaction (epPCR) and in vitro recombination of highly homologous sequences. For oligonucleotide-directed mutagenesis, we derive equations to estimate how complete a given library is expected to be and also to predict the size of library required to give a fixed probability of being 100% complete. We describe the statistical bases for computer programs which estimate the number of distinct variants represented in epPCR and shuffled libraries, dubbed PEDEL and DRIVeR, respectively. These programs allow the user to calculate (rather than guess) the ...

2003-06-01

43

Matrix-assisted laser desorption fourier transform mass spectrometry for biological compounds  

Science.gov (United States)

The recent development of matrix-assisted UV laser desorption (LD) mass spectrometry has made possible the ionization and detection of extremely large molecules (with molecular weights exceeding 100,000 Daltons). This technique has generated enormous interest in the biological community for the direct examination of large peptides and oligonucleotides. Although this matrix-assisted ionization method has been developed and used almost exclusively with time-of-flight (TOF) mass spectrometers, research is currently in progress to demonstrate this technique with trapped ion mass spectrometers, such as Fourier transform ion cyclotron resonance mass spectrometry (FTMS). The potential capabilities of FTMS for wide mass range, high resolution measurement, and ion trapping experiments suggest that this instrumental technique should be useful for the detailed structural characterization of large ions generated by the matrix-assisted technique. We have recently demonstrated ...

1990-01-01

44

Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9  

British Library Electronic Table of Contents (United Kingdom)

Abstract TLR9 recognizes unmethylated CpG-containing DNA commonly found in bacteria. Synthetic oligonucleotides containing CpG-motifs (CpG ODNs) recapitulate the activation of TLR9 by microbial DNA, whereas inversion of the CG dinucleotide within the CpG motif to GC (GpC ODNs) renders such ODNs inactive. This difference cannot be attributed to binding of ODNs to the full-length TLR9 ectodomain, as both CpG and GpC ODNs bind comparably. Activation of murine TLR9 requires cleavage into an active C-terminal fragment, which binds CpG robustly. We therefore compared the ability of CpG and GpC ODNs to bind to full-length and C-terminal TLR9, and their impact on the cleavage of TLR9. We found that CpG binds better to C-terminal TLR9 when compared with GpC, despite comparably low binding of both O...

2011-01-01

45

Locked nucleic acid (LNA): High affinity targeting of RNA for diagnostics and therapeutics  

DEFF Research Database (Denmark)

Locked nucleic acid (LNA) is a nucleic acid analogue containing one or more LNA nucleotide monomers with a bicyclic furanose unit locked in an RNA mimicking sugar conformation. This conformational restriction results in unprecedented hybridization affinity towards complementary single stranded RNA and thus, makes LNA uniquely suited for mimicking RNA structures and sequence specific targeting of RNA in vitro or in vivo. The focus of this paper is on LNAantisense, LNA-modified siRNA (siLNA), and detection and analysis of microRNAs by LNA-modified oligonucleotide probes.

2005-01-01

46

Source tracking of Escherichia coli by 16S-23S intergenic spacer region denaturing gradient gel electrophoresis (DGGE) of the rrnB ribosomal operon.  

Science.gov (United States)

This research validates a novel approach for source tracking based on denaturing gradient gel electrophoresis (DGGE) analysis of DNA extracted from Escherichia coli isolates. Escherichia coli from different animal sources and from river samples upstream from, at, and downstream of a combined sewer overflow were subjected to DGGE to determine sequence variations within the 16S-23S intergenic spacer region (ISR) of the rrnB ribosomal operon. The ISR was analyzed to determine if E. coli isolates from various animal sources could be differentiated from each other. DNA isolated from the E. coli animal sources was PCR amplified to isolate the rrnB operon. To prevent amplification of all 7 E. coli ribosomal operons by PCR amplification using universal primers, sequence-specific primers were utilized for the rrnB operon. Another primer set was then used to prepare samples of the 16S-23S ISR for DGGE. Comparison ...

2007-10-01

47

Transient suppression of MLH1 allows effective single-nucleotide substitution by single-stranded DNA oligonucleotides  

British Library Electronic Table of Contents (United Kingdom)

Short synthetic single-stranded oligodeoxyribonucleotides (ssODNs) can be used to introduce subtle modifications into the genome of mouse embryonic stem cells (ESCs). We have previously shown that effective application of ssODN-mediated gene targeting in ESC requires (transient) suppression of DNA mismatch repair (MMR). However, whereas transient down-regulation of the mismatch recognition protein MSH2 allowed substitution of 3 or 4 nucleotides, 1 or 2 nucleotide substitutions were still suppressed. We now demonstrate that single- or dinucleotide substitution can effectively be achieved by transient down-regulation of the downstream MMR protein MLH1. By exploiting highly specific real-time PCR, we demonstrate the feasibility of substituting a single basepair in a non-selectable gene. Howev...

2011-01-01

48

Isolation and mapping of a polymorphic DNA sequence (pEFD64. 2) on chromosome 3 (D3S46)  

Energy Technology Data Exchange (ETDEWEB)

A 4.0 kb TaqI fragment of cosmid EFD64 isolated by a HBV-3 oligonucleotide was subcloned into AccI site of pUC18. MspI identifies 5 allelic VNTR polymorphism with bands between 2.6 and 4.6 kb. RsaI, TaqI, EcoRI, BamHI, HindIII and PvuII identify the same VNTR polymorphism. With RsaI, 80% heterozygosity were observed in 80 unrelated Caucasians. This RFLP pEFD64.2 has been assigned to chromosome 3 by linkage analysis with loci (APOD) known to span this chromosome. Co-dominant segregation for the RsaI RFLP was observed in 40 three generation families. RFLPs were observed under the normal hybridization and washing condition.

1988-10-11

49

Species in the genus Turritopsis (Cnidaria, Hydrozoa): a molecular evaluation:  

British Library Electronic Table of Contents (United Kingdom)

Abstract Mitochondrial ribosomal gene sequences were used to investigate the status of several populations of hydromedusae belonging to the genus Turritopsis (family Oceaniidae). Several nominal species have been described for this genus, but most of them had been synonymized and attributed to one cosmopolitan species, Turritopsis nutricula. A recent revision based on morphological and reproductive characters, however, has shown that many different populations can be distinguished and that several of the nominal Turritopsis species are likely valid biological species. Our investigation using molecular sequence data of 16S mitochondrial gene confirms these results. The Mediterranean Turritopsis must be attributed to Turritopsis dohrnii and the Turritopsis of New Zealand must be referred to ...

2007-01-01

50

Molecular phylogenetic relationships and phenotypic diversity in miniaturized toadlets, genus Brachycephalus (Amphibia: Anura: Brachycephalidae)  

British Library Electronic Table of Contents (United Kingdom)

Toadlets of the genus Brachycephalus are endemic to the Atlantic rainforests of southeastern and southern Brazil. The 14 species currently described have snout-vent lengths less than 18mm and are thought to have evolved through miniaturization: an evolutionary process leading to an extremely small adult body size. Here, we present the first comprehensive phylogenetic analysis for Brachycephalus, using a multilocus approach based on two nuclear (Rag-1 and Tyr) and three mitochondrial (Cyt b, 12S, and 16S rRNA) gene regions. Phylogenetic relationships were inferred using a partitioned Bayesian analysis of concatenated sequences and the hierarchical Bayesian method (BEST) that estimates species trees based on the multispecies coalescent model. Individual gene trees showed conflict and also va...

2011-01-01

51

Methanogen community structure in the rumens of farmed sheep, cattle and red deer fed different diets  

British Library Electronic Table of Contents (United Kingdom)

Abstract Development of inhibitors and vaccines that mitigate rumen-derived methane by targeting methanogens relies on knowledge of the methanogens present. We investigated the composition of archaeal communities in the rumens of farmed sheep (Ovis aries), cattle (Bos taurus) and red deer (Cervus elaphus) using denaturing gradient gel electrophoresis (DGGE) to generate fingerprints of archaeal 16S rRNA genes. The total archaeal communities were relatively constant across species and diets, and were less variable and less diverse than bacterial communities. There were diet- and ruminant-species-based differences in archaeal community structure, but the same dominant archaea were present in all rumens. These were members of three coherent clades: species related to Methanobrevibacter ruminan...

2011-01-01

52

Functional adaptation of microbial communities from jet fuel-contaminated soil under bioremediation treatment: simulation of pollutant rebound  

British Library Electronic Table of Contents (United Kingdom)

Abstract To investigate the link between the functionality and the diversity of microbial communities under strong selective pressure from pollutants, two types of mesocosms that simulate natural attenuation and phytoremediation were generated using soil from a site highly contaminated with jet fuel and under air-sparging treatment. An increase in the petroleum hydrocarbon concentration from 4900 to 18-500-mg-kg-1-dw soil simulated a pollutant rebound (postremediation pollutant reversal due to residual contamination). Analysis of soil bacterial communities by denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments showed stronger changes and selection for a phylogenetically diverse microbial population in the mesocosms with pollutant-tolerant willow trees. Enumerat...

2011-01-01

53

Decolorization of textile azo dyes by newly isolated halophilic and halotolerant bacteria  

British Library Electronic Table of Contents (United Kingdom)

Studies were carried out on the decolorization of textile azo dyes by newly isolated halophilic and halotolerant bacteria. Among the 27 strains of halophilic and halotolerant bacteria isolated from effluents of textile industries, three showed remarkable ability in decolorizing the widely utilized azo dyes. Phenotypic characterization and phylogenetic analysis based on 16S rDNA sequence comparisons indicate that these strains belonged to the genus Halomonas. The three strains were able to decolorize azo dyes in a wide range of NaCl concentration (up to 20%w/v), temperature (25-40degreeC), and pH (5-11) after 4 days of incubation in static culture. They could decolorize the mixture of dyes as well as pure dyes. These strains also readily grew in and decolorized the high concentrations of dy...

2007-01-01

54

Clostridiaceae and Enterobacteriaceae as active fermenters in earthworm gut content  

British Library Electronic Table of Contents (United Kingdom)

The earthworm gut provides ideal in situ conditions for ingested heterotrophic soil bacteria capable of anaerobiosis. High amounts of mucus- and plant-derived saccharides such as glucose are abundant in the earthworm alimentary canal, and high concentrations of molecular hydrogen (H2) and organic acids in the alimentary canal are indicative of ongoing fermentations. Thus, the central objective of this study was to resolve potential links between fermentations and active fermenters in gut content of the anecic earthworm Lumbricus terrestris by 16S ribosomal RNA (rRNA)-based stable isotope probing, with [13C]glucose as a model substrate. Glucose consumption in anoxic gut content microcosms was rapid and yielded soluble organic compounds (acetate, butyrate, formate, lactate, propionate, succi...

2011-01-01

55

Chitinolytic enzyme production and genetic improvement of a new isolate belonging to Streptomyces anulatus  

British Library Electronic Table of Contents (United Kingdom)

Thirty bacterial isolates were obtained from different sources and sites at Jeddah, Saudi Arabia, on chitin agar medium; 9 of the 30 isolates were cultured in liquid medium containing chitin as sole carbon and nitrogen sources. Isolate SM21, which was isolated from shrimp shells, showed the best growth and chitinase production in liquid medium. According to its morphological, physiological and biochemical characteristics, SM21 belongs to the genus Streptomyces and was identified as Streptomyces anulatus SM21. Identification was confirmed using 16S rDNA analysis. The chitinase enzyme was precipitated with 80% NH4SO4 and purified using DEAE-cellulose ion exchange chromatography followed by Sephadex G-100 gel filtration. The molecular weight determined using sodium dodecyl sulfate polyacrylam...

2011-01-01

56

Transcription regulation of the vegf gene by the BMP/Smad pathway in the angioblast of zebrafish embryos  

Science.gov (United States)

Vascular endothelial growth factor (VEGF) is a mitogen that is critically involved in vasculogenesis, angiogenesis, and hematopoiesis. However, what and how transcription factors participate in the regulation of vegf gene expression are not fully understood. Here we report the cloning and sequencing of the zebrafish vegf promoter which revealed that the promoter contains a number of bone morphogenetic protein (BMP)-activated Smad binding elements (SBE), implicating Smad1 and Smad5 in the regulation of BMP-induced expression of vegf. Electrophoretic mobility shift assays of adding recombinant Smad proteins to the SBE-containing DNA oligonucleotides that represent portions of zebrafish vegf promoter resulted in mobility shift of the oligonucleotides. These changes demonstrate potential interactions between Smad1/5 and the vegf promoter. Reporter activity assays using the wild-type or SBE-deleted vegf promoters to drive the luciferase reporter ...

2005-04-01

57

Molecular cloning, characterization, and expression of human ADP-ribosylation factors: Two guanine nucleotide-dependent activators of cholera toxin  

International Nuclear Information System (INIS)

ADP-ribosylation factors (ARFs) are small guanine nucleotide-binding proteins that enhance the enzymatic activities of cholera toxin. Two ARF cDNAs, ARF1 and ARF3, were cloned from a human cerebellum library. Based on deduced amino acid sequences and patterns of hybridization of cDNA and oligonucleotide probes with mammalian brain poly(A)"+ RNA, human ARF1 is the homologue of bovine ARF1. Human ARF3, which differs from bovine ARF1 and bovine ARF2, appears to represent a newly identified third type of ARF. Hybridization patterns of human ARF cDNA and clone-specific oligonucleotides with poly(A)"+ RNA are consistent with the presence of at least two, and perhaps four, separate ARF messages in human brain. In vitro translation of ARF1, ARF2, and ARF3 produced proteins that behaved, by SDS/PAGE, similar to a purified soluble brain ARF. Deduced amino acid sequences of human ARF1 and ARF3 contain regions, similar to those in other G proteins, that ...

1989-01-01

58

Genetic heterogeneity in type 1 Gaucher disease: Multiple genotypes in Ashkenazic and non-Ashkenazic individuals  

International Nuclear Information System (INIS)

Nucleotide sequence analysis of a genomic clone from an Ashkenazic Jewish patient with type 1 Gaucher disease revealed a single-base mutation (adenosine to guanosine transition) in exon 9 of the glucocerebrosidase gene. This change results in the amino acid substitution of serine for asparagine. Transient expression studies following oligonucleotide-directed mutagenesis of the normal cDNA confirmed that the mutation results in loss of glucocerebrosidase activity. Allele-specific hybridization with oligonucleotide probes demonstrated that this mutation was found exclusively in type 1 phenotype. None of the 6 type 2 patients, 11 type 3 patients, or 12 normal controls had this allele. In contrast, 15 of 24 type 1 patients had one allele with this mutation, and 3 others were homozygous for the mutation. Furthermore, some of the Ashkenazic Jewish type 1 patients had only one allele with this mutation, suggesting that even in this population there is ...

59

Empirical Evaluation of a New Method for Calculating Signal to Noise Ratio (SNR) for Microarray Data Analysis  

Energy Technology Data Exchange (ETDEWEB)

Signal-to-noise-ratio (SNR) thresholds for microarray data analysis were experimentally determined with an oligonucleotide array that contained perfect match (PM) and mismatch (MM) probes based upon four genes from Shewanella oneidensis MR-1. A new SNR calculation, called signal to both standard deviations ratio (SSDR) was developed, and evaluated along with other two methods, signal to standard deviation ratio (SSR), and signal to background ratio (SBR). At a low stringency, the thresholds of SSR, SBR, and SSDR were 2.5, 1.60 and 0.80 with oligonucleotide and PCR amplicon as target templates, and 2.0, 1.60 and 0.70 with genomic DNA as target templates. Slightly higher thresholds were obtained at the high stringency condition. The thresholds of SSR and SSDR decreased with an increase in the complexity of targets (e.g., target types), and the presence of background DNA, and a decrease in the composition of targets, while SBR remained unchanged ...

2008-03-06

60

Monovalent cation-induced structure of telomeric DNA: The G-quartet model  

Energy Technology Data Exchange (ETDEWEB)

We have investigated the structures formed by oligonucleotides composed of two or four repeats of the telomeric sequences from Oxytricha and Tetrahymena. The Oxytricha four-repeat molecule (d(T4G4)4 = Oxy-4) forms structures with increased electrophoretic mobility in nondenaturing gels containing Na+, K+, or Cs+, but not in gels containing Li+ or no added salt. Formation of the folded structure results in protection of a set of dG's from methylation by dimethyl sulfate. Efficient UV-induced cross-links are observed in Oxy-4 and the related sequence from Tetrahymena (d(T2G4)4 = Tet-4), and join thymidine residues in different repeats. Models proposed to account for these data involve G-quartets, hydrogen-bonded structures formed from four guanosine residues in a square-planar array. We propose that the G-quartet structure must be dealt with in vivo by the telomere replication machinery.

1989-12-01

 
 
 
 
61

Identification of three related human GRO genes encoding cytokine functions  

International Nuclear Information System (INIS)

The product of the human GRO gene is a cytokine with inflammatory and growth-regulatory properties; GRO is also called MGSA for melanoma growth-stimulatory activity. The authors have identified two additional genes, GRO#beta# and GRO#gamma#, that share 90% and 86% identity at the deduced amino acid level with the original GRO#alpha# isolate. One amino acid substitution of proline in GRO#alpha# by leucine in GRO#beta# and GRO#gamma# leads to a large predicted change in protein conformation. Significant differences also exist in the 3' untranslated region, including different numbers of ATTTA repeats associated with mRNA instability. A 122-base-pair region in the 3' region is conserved among the three GRO genes, and a part of it is also conserved in the Chinese hamster genome, suggesting a role in regulation. DNA hybridization with oligonucleotide probes and partial sequence analysis of the genomic clones confirm that the three forms are derived from related but ...

62

Differentially expressed genes in hypothalamus in relation to genomic regions under selection in two chicken lines resulting from divergent selection for high or low body weight  

British Library Electronic Table of Contents (United Kingdom)

Long-term divergent selection for low or high body weight from the same founder population has generated two extremely divergent lines of chickens, the high- (HWS) and low-weight (LWS) selected lines. At selection age (56?days), the lines differ by more than nine times in body weight. The HWS line chickens are compulsive feeders, whereas in the LWS line, some individuals are anorexic and others have very low appetite. Previous studies have implicated the central nervous system and particularly the hypothalamus in these behavioural differences. Here, we compared the mRNA expression in hypothalamus tissue from chickens on day?4 post-hatch using oligonucleotide arrays and found that the divergent selection had resulted in minor but multiple expression differences. Differentially expressed gen...

2011-01-01

63

DNA alterations photosensitized by tetracycline and some of its derivatives  

Energy Technology Data Exchange (ETDEWEB)

Bacteriophage M13 mp10 DNA were irradiated with near-UV light in the presence of tetracycline derivatives and primed with synthetic oligonucleotide to be used for DNA synthesis using Escherichia coli DNA polymerase. Chain terminations were observed by denaturing polyacrylamide gel electrophoresis and mapped precisely. All the synthesis stops occurred before or at the level of guanine residues, showing that the photoreaction mediated by tetracycline derivatives led to a preferential alteration of guanine residues. These lesions were demonstrated to be induced in DNA through a pathway involving singlet oxygen. Tetracycline derivatives also photoinduced the breakage of the DNA sugar-phosphate backbone monitored by the conversion of supercoiled phi X174 DNA to a relaxed form. This lesion was shown to be initiated by hydroxyl radicals. The production of this free radical has been confirmed by electron paramagnetic resonance (EPR) spin trapping experiments using ...

1986-06-01

64

Cloning and sequencing of cDNA encoding human DNA topoisomerase II and localization of the gene to chromosome region 17q21-22  

Energy Technology Data Exchange (ETDEWEB)

Two overlapping cDNA clones encoding human DNA topoisomerase II were identified by two independent methods. In one, a human cDNA library in phage {lambda} was screened by hybridization with a mixed oligonucleotide probe encoding a stretch of seven amino acids found in yeast and Drosophila DNA topoisomerase II; in the other, a different human cDNA library in a {lambda}gt11 expression vector was screened for the expression of antigenic determinants that are recognized by rabbit antibodies specific to human DNA topoisomerase II. The entire coding sequences of the human DNA topoisomerase II gene were determined from these and several additional clones, identified through the use of the cloned human TOP2 gene sequences as probes. Hybridization between the cloned sequences and mRNA and genomic DNA indicates that the human enzyme is encoded by a single-copy gene. The location of the gene was mapped to chromosome 17q21-22 by in situ hybridization of a cloned fragment to ...

1988-10-01

65

Cellular uptake of Aib-containing amphipathic helix peptide.  

Science.gov (United States)

Cell-penetrating peptides (CPPs) are useful tools for the delivery of hydrophilic bioactive molecules, such as peptides, proteins, and oligonucleotides, across the cell membrane. To realize the delivery of therapeutic macromolecules by CPPs, the CPPs are required to show resistance to protease and no cytotoxicity. In order to produce potent non-toxic and protease-resistant CPPs with high cellular uptake, we designed an amphipathic helix peptide using ?-aminoisobutyric acid (Aib, U) and named it MAP(Aib). In the MAP(Aib) molecule, five Aib residues are aligned on the hydrophobic face of the helix and five lysine (K) residues are aligned on the hydrophilic face. MAP(Aib) showed potent resistance to trypsin and pronase compared with MAP, an amphipathic helix peptide formed by usual amino acids. Fluorescein-labeled MAP(Aib) efficiently traversed the A549 cell membrane, diffusing into the cytoplasm and slightly into the nucleus without exerting any cytotoxicity. In ...

2011-08-11

66

Biophysical analysis of DNA modified by 1,2-diaminocyclohexane platinum(II) complexes  

Energy Technology Data Exchange (ETDEWEB)

Modification of DNA and double-stranded deoxyoligonucleotides with antitumor 1,2-diamino-cyclohexanedinitroplatinum(II) (Pt-dach) complexes was investigated with the aid of physico-chemical methods and chemical probes of nucleic acid conformation. The three Pt-dach complexes were used which differed in isomeric forms of the dach non-leaving ligand-Pt(1R,2R-dach), Pt(1S,2S-dach) and Pt(1R,2S-dach) complexes. The latter complex has lower antitumor activity than the other two Pt-dach complexes. Pt(1R,2S-dach) complex exhibits the slowest kinetics of its binding to DNA and of the conversion of monofunctional binding to bifunctional lesions. The anomalously slow electrophoretic mobility of multimers of the platinated and ligated oligomers suggests that bifunctional binding of Pt-dach complexes to a d(GG) site within double-stranded oligonucleotides induces bending of the oligomer. It is proposed that different effects of the Pt-dach complexes on DNA observed in this ...

1992-01-25

67

The mitochondrial genome of the entomophagous endoparasite Xenosvesparum (Insecta: Strepsiptera)  

Energy Technology Data Exchange (ETDEWEB)

In this study, the nearly complete sequence (14,519 bp) of the mitochondrial DNA (mtDNA) of the entomophagous endoparasite Xenos vesparum (Insecta: Strepsiptera) is described. All protein coding genes (PCGs) are in the arrangement known to be ancestral for insects, but three tRNA genes (trnA, trnS(gcu), and trnL(uag)) have transposed to derived positions and there are three tandem copies of trnH, each of which is potentially functional. All of these rearrangements except for that of trnL(uag) is within the short span between nad3 and nad4 and there are numerous blocks of unassignable sequence in this region, perhaps as remnants of larger scale predisposing rearrangements. X. vesparum mtDNA nucleotide composition is strongly biased toward As and Ts, as is typical for insect mtDNAs. There is also significant strand skew in the distribution of these nucleotides, with the J-strand being richer in A than T and in C than G, and the N-strand showing an opposite skew for complementary pairs of ...

2005-12-01

68

Report on a survey in fiscal 1999. Survey on industrial utilization of microorganism reaction mechanisms under anaerobic condition; 1999 nendo kenki jokenka ni okeru biseibutsu hanno kiko no kogyoteki riyo ni kansuru chosa hokokusho  

Energy Technology Data Exchange (ETDEWEB)

Industrial utilization of reaction mechanisms of microorganisms under anaerobic condition permits structuring energy saving type production processes. The present survey has investigated features of new microorganisms under anaerobic condition and the status of researches thereon inside and outside the country, and discussed their future applications. Chapter 1 compares anaerobic microorganisms and functions of microorganism under anaerobic condition with those aerobic to describe their general features, and describes the purpose of this survey and the summary of the investigations. Chapter 2 surveys the current status of technologies to utilize microorganisms under anaerobic condition. Chapter 3 outlines metabolic characteristics of the anaerobic microorganisms, and extracts functions effective for material production by different anaerobic microorganisms to describe their applicability. Chapter 4 evaluates the system classification for the anaerobic microorganisms utilizing the basic ...

2000-03-01

69

Isolation and Identification of Achromobacter sp. DN-06 and Evaluation of Its Pyridine Degradation Kinetics  

British Library Electronic Table of Contents (United Kingdom)

We report the isolation of a new bacterium species (named as DN-06) that degrades pyridine, a model compound containing both carbon and nitrogen, from the aerobic activated sludge in a coking wastewater treatment plant. DN-06 was identified as Achromobacter sp. using 16S rDNA sequence analysis. In batch culture, more than 95% of pyridine (500?mg/L) was degraded within 18?h by DN-06 grown at 35?C and pH?8 with agitation at 170?rpm. Degradation experiments of pyridine at different initial concentrations (50?4,300?mg/L) revealed that pyridine was an inhibitory substrate, and that neither yield coefficient Y nor endogenous decay coefficient K d was a constant. The values of Y and K d were 0.55?0.74 and 0.0032?0.0057?h?1, respectively. Five kinetic models (Haldane, Yano, Aiba, Webb, and Monod) ...

2011-01-01

70

Identification of culturable and originally non-culturable endophytic bacteria isolated from shoot tip cultures of banana cv. Grand Naine  

British Library Electronic Table of Contents (United Kingdom)

In this article we describe the identification of endophytic bacteria belonging to three groups isolated from shoot tip cultures of banana cv. Grand Naine in a recent study (Thomas et?al. 2008) based on partial 16S rRNA gene sequence homology analysis. The first group included banana stocks that displayed obvious colony growth on MS based tissue culture medium during the first in?vitro passage. The second group constituted stocks that were tissue index-negative for cultivable bacteria initially but turned index-positive after a few to several (4?8) in?vitro passages while the third group formed one sub-stock that turned index-positive after about 18 passages. The organisms belonged to about 20 different genera comprising of ?, ?, ?-proteobacteria, Gram-positive firmicutes and actinobacteri...

2008-01-01

71

Evidence of minimal methanogenic numbers and activity in sediments collected from the JAPEX/JNOC/GSC et al. Mallik 5L-38 gas hydrate production research well  

Energy Technology Data Exchange (ETDEWEB)

Gene analysis was used to determined the presence, abundance and phylogenetic affiliation of methanogens that exist in gas-hydrate-bearing sediment samples obtained from 23 drill cores from the JAPEX/JNOC/GSC et al. Mallik 5L-38 gas hydrate research well. Rates of methane production were examined using sediment-inoculated enrichments containing {sup 14}C-labeled carbon substrates, carbon dioxide and acetate. Archaeal 16S rDNA was only detected in 6 of the samples, resulting in 8 sequences with relationships to the Miscellaneous Crenarchaeotic Group (7 clones) and the Subsurface Euryarchaeotic Group (1 clone). The single Euryarchaeota sequence did not appear to be related to methanogens. Subsamples from the cores showed variable results upon DNA extraction and amplification. Methanogenic Coenzyme M (CoM) was detected in 13 of the 20 cores, but methanogenic methyl CoM reductase genes were not amplified from the samples when using a sensitive ...

2005-07-01

72

Biological control of take-all in wheat by endophytic Bacillus subtilis E1R-j and potential mode of action  

British Library Electronic Table of Contents (United Kingdom)

The bacterial strain E1R-j, isolated as an endophyte from wheat roots, exhibited high antifungal activity to Gaeumannomyces graminis var. tritici (Ggt). Strain E1R-j was identified as Bacillus subtilis based on morphological, physiological and biochemical methods as well as on 16S rDNA analysis. This strain inhibited mycelium growth in vitro of numerous plant pathogenic fungi, especially of Ggt, Coniothyrium diplodiella, Phomopsis sp. and Sclerotinia sclerotiorum. In greenhouse experiments, soil drenches with cell densities of 106, 109 and 1012CFU ml-1 E1R-j reduced significantly take-all disease, caused by Ggt, in wheat seedling by 62.6%, 68.6% and 70.7%, respectively, compared to the inoculated control, 4weeks after sowing. Growth parameters such as lengths and fresh weights of roots and...

2009-01-01

73

Resistance to powdery mildew in Spanish barley landraces is controlled by different sets of quantitative trait loci.  

Science.gov (United States)

Twenty-two landrace-derived inbred lines from the Spanish Barley Core Collection (SBCC) were found to display high levels of resistance to a panel of 27 isolates of the fungus Blumeria graminis that exhibit a wide variety of virulences. Among these lines, SBCC145 showed high overall resistance and a distinctive spectrum of resistance compared with the other lines. Against this background, the main goal of the present work was to investigate the genetic basis underlying such resistance using a doubled haploid population derived from a cross between SBCC145 and the elite spring cultivar Beatrix. The population was genotyped with the 1,536-SNP Illumina GoldenGate Oligonucleotide Pool Assay (Barley OPA-1 or BOPA1 for short), whereas phenotypic analysis was performed using two B. graminis isolates. A major quantitative trait locus (QTL) for resistance to both isolates was identified on the long arm of chromosome 6H (6HL) and accounted for ca. 60% of the phenotypic ...

2011-07-08

74

Investigation of genomic instability by assay of DNA fingerprint from the offspring of male mice exposed to chronic low-level #gamma#-radiation  

International Nuclear Information System (INIS)

By polymerase chain reaction with arbitrary primer (AP-PCR), the possibility of transmission of genome instability to somatic cells of the offspring (F_1 generation) from male parents of mice exposed to chronic low-dose #gamma#-radiation was studied. Male mice 15 days after exposure to 10-50 cGy were mated with unirradiated females. Biopsies were taken from tale tips of two month-old mice progeny for DNA separation. Primer in the AP-PCR was 20-mer oligonucleotide flanking the micro-satellite locus Atplb2 on chromosome 11 of the mouse. Comparative analysis of individual fingerprints of AP-PCR products on DNA-templates from the offspring of irradiated and unirradiated male mice revealed an increased variability of micro-satellite-associated sequences in the genome of the offspring of males exposed to 25 and 50 cGy. DNA-fingerprints of the offspring of male mice exposed to chronic irradiation doses 10 and 25 cGy. 15 days before fertilization (at the post-meiotic stage ...

2000-11-20

75

Detecting and Genotyping Escherichia coli O157:H7 using multiplexed PCR and nucleic acid microarrays  

Energy Technology Data Exchange (ETDEWEB)

Rapid detection and characterization of food borne pathogens such as Escherichia coli O157:H7 is crucial for epidemiological investigations and food safety surveillance. As an alternative to conventional technologies, we examined the sensitivity and specificity of nucleic acid microarrays for detecting and genotyping E. coli O157:H7. The array was composed of oligonucleotide probes (25-30 mer) complementary to four virulence loci (intimin, Shiga-like toxins I and II, and hemolysin A). Target DNA was amplified from whole cells or from purified DNA via single or multiplexed polymerase chain reaction (PCR), and PCR products were hybridized to the array without further modification or purification. The array was 32-fold more sensitive than gel electrophoresis and capable of detecting amplification products from < 1 cell equivalent of genomic DNA (1 fg). Immunomagnetic capture, PCR and a microarray were subsequently used to detect 55 CFU ml-1 (E. coli O157:H7) ...

2000-12-01

76

Cloning of human basic A1, a distinct 59-kDa dystrophin-associated protein encoded on chromosome 8q23-24  

Energy Technology Data Exchange (ETDEWEB)

Duchenne and Becker muscular dystrophies are caused by defects of dystrophin, which forms a part of the membrane cytoskeleton of specialized cells such as muscle. It has been previously shown that the dystrophin-associated protein A1 (59-kDa DAP) is actually a heterogeneous group of phosphorylated proteins consisting of an acidic ({alpha}-A1) and a distinct basic ({beta}-A1) component. Partial peptide sequence of the A1 complex purified from rabbit muscle permitted the design of oligonucleotide probes that were used to isolate a cDNA for one human isoform of A1. This cDNA encodes a basic A1 isoform that is distinct from the recently described syntrophins in Torpedo and mouse and is expressed in many tissues with at least five distinct mRNA species of 5.9, 4.8, 4.3, 3.1, and 1.5 kb. A comparison of the human cDNA sequence with the GenBank expressed sequence tag (EST) data base has identified a relative from human skeletal muscle, EST25263, which is probably a human ...

1994-05-10

77

Alteration of nucleoside diphosphate binding specificity of E. coli elongation factor Tu (EF-Tu) by single amino acid substitution at position 138  

International Nuclear Information System (INIS)

A single amino acid substitution (Asp #-># Asn) at position 138 of E. coli EF-Tu was induced in the tufA gene by an M13 phage oligonucleotide site-directed mutagenesis protocol. The mutated tufA gene was then subcloned in a plasmid vector and expressed in maxicells. The properties of ["3"5S]methionine labelled mutant and wild type EF-Tu's were compared by in vitro assays. Mutant and wild-type EF-Tu's bound EF-Ts with approximately equal affinities. The 138-Asn mutation greatly reduced the protein's affinity for GDP; however, this mutation dramatically increased the proteins affinity for XDP. The mutant protein forms a stable complex with phe-tRNA and XTP, which binds to ribosomes; whereas, it does not form a complex with phe-tRNA and GTP. These results suggest that in EF-Tu x NDP complexes amino acid residue 138 must interact with the substituent on C-2 of the purine ring. Thus in wild-type EF-Tu Asp-138 would H-bond to 2-NH_2 of GDP, and in the mutant EF-Tu ...

1987-06-07

78

3-Methyl-3-deazaadenine, a stable isostere of N3-methyl-adenine, is efficiently bypassed by replication in vivo and by transcription in vitro.  

Science.gov (United States)

The goal of the present work was to determine the impact of N3-methyladenine (3-mA), an important lesion generated by many environmental agents and anticancer drugs, on in vivo DNA replication and in vitro RNA transcription. Due to 3-mA chemical instability, the stable isostere 3-methyl-3-deazaadenine (3-m-c(3)A) was site specifically positioned into an oligodeoxynucleotide. The oligomer was, then incorporated into a vector system that is rapidly converted to ssDNA inside yeast cells and requires DNA replication opposite the lesion for plasmid clonal selection. For control purposes, an adenine or a stable apurinic/apyrimidinic (AP)-lesion was placed at the same site. The presence of each lesion in the oligonucleotide was confirmed by MALDI-TOF analysis. Plasmids were then transfected into yeast cells. While the AP-site dramatically reduced plasmid replication in all strains, the 3-m-c(3)A had a slight effect in the rad30 background which significantly increased ...

2011-06-14

79

Thermochemistry of mixed explosives  

Energy Technology Data Exchange (ETDEWEB)

In order to predict thermal hazards of high-energy materials, accurate kinetics constants must be determined. Predictions of thermal hazards for mixtures of high-energy materials require measurements on the mixtures, because interactions among components are common. A differential-scanning calorimeter (DSC) can be used to observe rate processes directly, and isothermal methods enable detection of mechanism changes. Rate-controlling processes will change as components of a mixture are depleted, and the correct depletion function must be identified for each specific stage of a complex process. A method for kinetics measurements on mixed explosives can be demonstrated with Composition B is an approximately 60/40 mixture of RDX and TNT, and is an important military explosive. Kinetics results indicate that the mator process is the decomposition of RDX in solution in TNT with a perturbation caused by interaction between the two components. It is concluded that a combination of chemical ...

1982-01-01

80

Single protein omission reconstitution studies of tetracycline binding to the 30S subunit of Escherichia coli ribosomes  

Energy Technology Data Exchange (ETDEWEB)

In previous work the authors showed that on photolysis of Escherichia coli ribosomes in the presence of ({sup 3}H)tetracycline (TC) the major protein labeled is S7, and they presented strong evidence that such labeling takes place from a high-affinity site related to the inhibitory action of TC. In this work they use single protein omission reconstitution (SPORE) experiments to identify those proteins that are important for high-affinity TC binding to the 30S subunit, as measured by both cosedimentation and filter binding assays. With respect to both sedimentation coefficients and relative Phe-tRNA{sup Phe} binding, the properties of the SPORE particles they obtain parallel very closely those measured earlier, with the exception of the SPORE particle lacking S13. A total of five proteins, S3, S7, S8, S14, and S19, are shown to be important for TC binding, with the largest effects seen on omission of proteins S7 and S14. Determination of the protein compositions of the corresponding ...

1990-06-05

 
 
 
 
81

Patterns of lineage diversification in the genus Naso (Acanthuridae).  

Science.gov (United States)

The evolutionary history of the reef fish genus Naso (F. Acanthuridae) was examined using a complete species-level molecular phylogeny of all recognized (19) species based on three loci (one nuclear ETS2 and two mitochondrial 16S, cyt b). This study demonstrates that distinct foraging modes and specialized body shapes arose independently at different times in the evolutionary history of the genus. Members of the subgenus Axinurus, characterized by a scombriform morphology, caudal fin structure and pelagic foraging mode, were consistently placed basal to the remaining Naso species, suggesting that pelagic foraging is plesiomorphic and benthic foraging derived in this genus. We used a genus-level phylogeny (nuclear marker, ETS2), which included several taxa from all other acanthurid genera, to obtain a range of age estimates for the most recent common ancestor of the genus Naso. These age estimates (range of 52-43.3 MY) were then used to estimate ...

2004-07-01

82

Microbiological Comparison of Core and Groundwater Samples Collected from a Fractured Basalt Aquifier with that of Dialysis Chamber Incubated in Situ  

Energy Technology Data Exchange (ETDEWEB)

Microorganisms associated with fractured basalt core were compared to those suspended in groundwater pumped from the same well in the eastern Snake River Plain Aquifer (Idaho, USA). Two wells influenced to different degrees by a mixed-waste plume in the fractured basalt aquifer were examined. In one well, an array of dialysis cells filled with either deionized water or crushed basalt was equilibrated to compare the microorganisms collected in this fashion with those from core and groundwater samples collected in a traditional manner from the same well. Analyses were performed to characterize these samples and to provide a basis for comparison. These included total cell counts by microscopy; total biomass by phospholipid fatty acid analysis; enumerations of viable aerobic heterotrophs, groups of putative aerobic co-metabolic TCE-degraders and aerobic H2-oxidizing bacteria; mineralization of 14C- labeled acetate; and enrichments for dissimilatory iron-reducing bacteria. The ...

2004-04-01

83

Microbial community analysis of ambient temperature anaerobic digesters  

Energy Technology Data Exchange (ETDEWEB)

This paper reported on a study in which designs for Chinese and Indian fixed-dome anaerobic digesters were modified in an effort to produce smaller and more affordable digesters. While these types of systems are common in tropical regions of developing countries, they have not been used in colder climates because of the low biogas yield during the winter months. Although there is evidence that sufficient biogas production can be maintained in colder temperatures through design and operational changes, there is a lack of knowledge about the seasonal changes in the composition of the microbial communities in ambient temperature digesters. More knowledge is needed to design and operate systems for maximum biogas yield in temperate climates. The purpose of this study was to cultivate a microbial community that maximizes biogas production at psychrophilic temperatures. The study was conducted on a 300 gallon experimental anaerobic digester on the campus of Ohio State University. ...

2010-07-01

84

Biogeochemistry of the compost bioreactor components of a composite acid mine drainage passive remediation system  

International Nuclear Information System (INIS)

The compost bioreactor ('anaerobic cell') components of three composite passive remediation systems constructed to treat acid mine drainage (AMD) at the former Wheal Jane tin mine, Cornwall, UK were studied over a period of 16 months. While there was some amelioration of the preprocessed AMD in each of the three compost bioreactors, as evidenced by pH increase and decrease in metal concentrations, only one of the cells showed effective removal of the two dominant heavy metals (iron and zinc) present. With two of the compost bioreactors, concentrations of soluble (ferrous) iron draining the cells were significantly greater than those entering the reactors, indicating that there was net mobilisation (by reductive dissolution) of colloidal and/or solid-phase ferric iron compounds within the cells. Soluble sulfide was also detected in waters draining all three compost bioreactors which was rapidly oxidised, in contrast to ferrous iron. Oxidation and hydrolysis of iron, together with ...

2005-02-01

85

ZZ MCJEF22NEA.BOLIB, MCNP Cross Section Library Based on JEF-2.2  

International Nuclear Information System (INIS)

1 - Description or function: Continuous energy cross-section data library for the Monte Carlo program MCNP based on the JEF-2.2 evaluated nuclear data library (ACE Format). Format: ACE Number of groups: Continuous energy Nuclides (107): H-1, H-2, He-4, Li-6, Li-7, Be-9, B-10, B-11, C-nat, N-14, N-15, O-16, O-17, F-19, Na-23, Mg-nat, Al-27, Si-nat, Cl-nat, Ti-nat, Cr-50, Cr-52, Cr-53, Cr-54, Mn-55, Fe-54, Fe-56, Fe-57, Fe-58, Co-59, Ni-58, Ni-60, Ni-61, Ni-62, Ni-64, Zr-90, Zr-91, Zr-92, Zr-94, Zr-96, Zr-nat, Nb-93, Mo-92, Mo-94, Mo-95, Mo-96, Mo-97, Mo-98, Mo-100, Mo-nat, Tc-99, Ru-101, Ru-102, Ru-104, Rh-103, Pd-105, Pd-107, Ag-109, I-129, Xe-131, Cs-133, Pr-141, Nd-143, Nd-145, Pm-147, Sm-147, Sm-149, Sm-150, Sm-151, Sm-152, Eu-153, Gd-154, Gd-155, Gd-156, Gd-157, Gd-158, Gd-160, Hf-174, Hf-176, Hf-177, Hf-178, Hf-179, Hf-180, Pb-nat, Bi-209, Th-232, U-234, U-235, U-236, U-238, Np-237, Pu-238, Pu-239, Pu-239bis, Pu-240, Pu-241, Pu-242, Am-241, Am-242, Am-242m, Am-243, Cm-242, Cm-243, ...

86

ZZ MCB63NEA.BOLIB, MCNP Cross Section Library Based on ENDF/B-VI Release 3  

International Nuclear Information System (INIS)

1 - Description of program or function: Continuous energy cross-section data library for the Monte Carlo program MCNP based on the ENDF/B-VI Release 3 evaluated nuclear data library (ACE Format). Format: ACE; Number of groups: Continuous energy; Nuclides (107): H-1, H-2, He-4, Li-6, Li-7, Be-9, B-10, B-11, C-nat, N-14, N-15, O-16, O-17, Na-23, Mg-nat, Al-27, Si-nat, Cl-nat, Ti-nat, Cr-50, Cr-52, Cr-53, Cr-54, Mn-55, Fe-54, Fe-56, Fe-57, Fe-58, Co-59, Ni-58, Ni-60, Ni-61, Ni-62, Ni-64, Zr-90, Zr-91, Zr-92, Zr-94, Zr-96, Zr-nat, Nb-93, Mo-94, Mo-95, Mo-96, Mo-97, Mo-nat, Tc-99, Ru-101, Ru-102, Ru-104, Rh-103, Pd-105, Pd-107, Ag-109, I-129, Xe-131, Cs-133, Pr-141, Nd-143, Nd-145, Pm-147, Sm-147, Sm-149, Sm-150, Sm-151, Sm-152, Eu-153, Gd-154, Gd-155, Gd-156, Gd-157, Gd-158, Gd-160, Hf-174, Hf-176, Hf-177, Hf-178, Hf-179, Hf-180, Hf-nat, Pb-206, Pb-207, Pb-208, Bi-209, Th-232,U-233, U-234, U-235, U-236, U-238, Np-237, Pu-238, Pu-239, Pu-240, Pu-241, Pu-242, Am-241, Am-242, Am-242m, Am-243, ...

87

WLUP3.0, 69 and 172 Group Cross Section Libraries for WIMS  

International Nuclear Information System (INIS)

Description or function: WLUP contains validated WIMS-D formatted cross section libraries in 69 and 172 energy group structures for nuclear reactor calculations. Materials from recently released evaluated nuclear data libraries are included. The NJOY nuclear data processing system was applied for generating the cross section files following the models and conventions built into the WIMS-D lattice code. The relevant features for the WIMS users are: - Energy group structures: 69 and 172 energy groups. - List of materials: WIMS ID, general information, source of data. - Cross sections: 69 and 172 group plots. - Resonance data: WIMS ID, temperature, background cross sections. - Goldstein-Cohen factors: Goldstein-Cohen lambda values. - Thermal scattering data: thermal scattering laws and P1 matrixes. - Fission spectrum: fission spectrum data. - Burnup data: burnup chains. - Fission product yields: fission yield tables. - Pseudo lumped fission product: Description of pseudo fission product. ...

88

ZZ KASHIL-E6, 175 N, 42 Gamma Groups Cross Sections in MATXS Format Based on ENDF/B-VI.5 for Shielding Applications  

International Nuclear Information System (INIS)

1 - Description of program or function: Format: MATXS; Number of groups: 175 neutron-, 42 photon-groups; 176 Nuclides: 1-H-1, 1-H-2, 1-H-3, 2-He-3, 2-He-4, 3-Li-6, 3-Li-7, 4-Be-9, 5-B-10, 5-B-11, 6-C- nat., 7-N-14, 7-N-15, 8-O-16, 8-O-17, 9-F-19, 11-Na-23, 12-Mg-nat., 13-Al-27, 14-Si-nat., 14-Si-28, 14-Si-29, 14-Si-30, 15-P-31, 16-S-32, 17-Cl-nat., 19-K-nat., 20-Ca-nat., 21-Sc-45, 22-Ti-nat., 23-V-nat., 24-Cr-50, 24-Cr-52, 24-Cr-53, 24-Cr-54, 25-Mn-25, 26-Fe-54, 26-Fe-56, 26-Fe-57, 26-Fe-58, 27-Co-59, 28-Ni-58, 28-Ni-60, 28-Ni-61, 28-Ni-62, 28-Ni-64, 29-Cu-63, 29-Cu-65, 31-Ga-nat., 39-Y-89, 40-Zr-nat., 40-Zr-90, 40-Zr-91, 40-Zr-92, 40-Zr-94, 40-Zr-96, 41-Nb-93, 42-Mo-nat., 46-Pd-102, 46-Pd-104, 46-Pd-105, 46-Pd-106, 46-Pd-108, 46-Pd-110, 47-Ag-107, 47-Ag-109, 48-Cd-106, 48-Cd-108, 48-Cd-110, 48-Cd-112, 48-Cd-113, 48-Cd-114, 48-Cd-116, 49-In-nat., 53-I-127, 54-Xe-124, 54-Xe-126, 54-Xe-128, 54-Xe-129, 54-Xe-130, 54-Xe-131, 54-Xe-132, 54-Xe-134, ...

89

ZZ DECAYREM/C, Decay Spectra Library for EXREM Calculation  

International Nuclear Information System (INIS)

Description of problem or function: Format: EXREM III; Nuclides: radioactive decay data on 252 Nuclides: 1H-3, 4Be-7, 6C-11, 6C-14, 7N-13, 8O-15, 9F-18, 11Na-22, 11Na-24, 12Mg-28, 13Al-28, 15P-32, 15P-33, 16S-35, 17Cl-36, 17Cl-38, 18A-37, 18A-39, 19K-40, 19K-42, 19K-43, 20Ca-45, 20Ca-47, 20Ca-49, 21Sc-46, 21Sc-47, 21Sc-49, 24Cr-51, 25Mn-52M, 25Mn-52, 25Mn-54, 26Fe-52, 26Fe-55, 26Fe-59, 27Co-56, 27Co-57, 27Co-58, 27Co-60, 28Ni-56, 28Ni-63, 29Cu-64, 30Zn-65, 30Zn-69M, 30Zn-69, 31Ga-67, 31Ga-68, 32Ge-77, 33As-76, 33As-77, 34Se-75, 35Br-80M, 35Br-80, 35Br-82, 35Br-83, 35Br-84, 36Kr-79, 36Kr-83M, 36Kr-85M, 36Kr-85, 36Kr-87, 36Kr-88, 37Rb-84, 37Rb-86, 37Rb-87, 37Rb-88, 37Rb-89, 37Rb-90M, 37Rb-90, 38Sr-85, 38Sr-87M, 38Sr-89, 38Sr-90, 38Sr-91, 38Sr-92, 38Sr-93, 39Y-87, 39Y-88, 39Y-90, 39Y-91M, 39Y-91, 39Y-92, 39Y-93, 40Zr-93, 41Nb-93M, 40Zr-95, 40Zr-97, 41Nb-95M, 41Nb-95, 41Nb-97M, 41Nb-97, 42Mo-99, 43Tc-99M, 43Tc-99, 44Ru-103, 44Ru-105, 44Ru-106, ...