[Differential expression of DTSsa4 Tc1-like transposons in closely related populations of Baikal ciscoes].

Science.gov (United States)

Bychenko, O S; Sukhanova, L V; Azhikina, T L; Sverdlov, E D

2009-01-01

Two representatives of Baikal ciscoes - lake cisco and omul - diverged from a common ancestor as recently as 10-20 thousand years ago. We have found an increasing expression level of DTSsa4 Tc1-like DNA transposons in cisco and omul brains. The mapping of the sequences of these transposons from Salmo salar and Danio rerio genomes has shown that in some cases, these transposons are located in the 5' and 3' regions, as well as in the promoter regions of various genes. Probably, Tc1-like transposons affect the activity of neighboring genes, providing the adaptive divergence of the cisco population.

Induction of transposon TN1 translocation under the action of different mutagens

International Nuclear Information System (INIS)

Kubanejshvili, M.G.; Smirnov, S.P.; Tarasov, V.A.

1983-01-01

Migration of ampicillin transposon Tn1 under normal conditions in Escherichia coli cells proceeds with low frequency (10 -4 transpositions for cell). The low transposition frequency is conditioned by the transposition repression, realized by the gene-repressor in transposon structure and, probably, by other regulating genes of the bacterium-host. E. coli cell treatment by physical and chemical mutagens resulted in induction of translocation of ampicillin transposon Tn1 from plasmid RP4 into other replicons. Mitomycin C and ultraviolet radiation produced stronger inducing effect as compared to nitroso-guanidine (NG). The effect of the given mutagens on transposition Tn1 correlated with their activating capacity with respect to inducible SOS-functions of E coli. The mutation of rec A didn't influence on spontaneous Tn1 transposition, but blocked completely the induction of transposition process under mutagen effect. The relationship of inducible transposition with SOS-functions in E. coli cells, controlled by recA and lexA genes, as well as the possible role of the process in genetic microorganism variability are discussed in the paper

Global mapping of transposon location.

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Abram Gabriel

2006-12-01

Full Text Available Transposable genetic elements are ubiquitous, yet their presence or absence at any given position within a genome can vary between individual cells, tissues, or strains. Transposable elements have profound impacts on host genomes by altering gene expression, assisting in genomic rearrangements, causing insertional mutations, and serving as sources of phenotypic variation. Characterizing a genome's full complement of transposons requires whole genome sequencing, precluding simple studies of the impact of transposition on interindividual variation. Here, we describe a global mapping approach for identifying transposon locations in any genome, using a combination of transposon-specific DNA extraction and microarray-based comparative hybridization analysis. We use this approach to map the repertoire of endogenous transposons in different laboratory strains of Saccharomyces cerevisiae and demonstrate that transposons are a source of extensive genomic variation. We also apply this method to mapping bacterial transposon insertion sites in a yeast genomic library. This unique whole genome view of transposon location will facilitate our exploration of transposon dynamics, as well as defining bases for individual differences and adaptive potential.

The rde-1 gene, RNA interference, and transposon silencing in C. elegans.

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Tabara, H; Sarkissian, M; Kelly, W G; Fleenor, J; Grishok, A; Timmons, L; Fire, A; Mello, C C

1999-10-15

Double-stranded (ds) RNA can induce sequence-specific inhibition of gene function in several organisms. However, both the mechanism and the physiological role of the interference process remain mysterious. In order to study the interference process, we have selected C. elegans mutants resistant to dsRNA-mediated interference (RNAi). Two loci, rde-1 and rde-4, are defined by mutants strongly resistant to RNAi but with no obvious defects in growth or development. We show that rde-1 is a member of the piwi/sting/argonaute/zwille/eIF2C gene family conserved from plants to vertebrates. Interestingly, several, but not all, RNAi-deficient strains exhibit mobilization of the endogenous transposons. We discuss implications for the mechanism of RNAi and the possibility that one natural function of RNAi is transposon silencing.

Microbial Fe (III) reduction and hydrogen production by a transposon-mutagenized strain of Pantoea agglomerans BH18

International Nuclear Information System (INIS)

Liu, Hongyan; Wang, Guangce

2015-01-01

Based on the transposon-mutagenized library of Pantoea agglomerans BH18, mutant screens were conducted to obtain the strain with the highest Fe (III) reduction and hydrogen production. Of these transposon-mutagenized mutants, the mutant strain TB230 was screened for high Fe (III)-reducing efficiency and hydrogen production. The PCR amplification and kanamycin resistance selection results indicated that the transposon insertion of the mutant strain TB230 was stable. Hydrogen production of the mutant strain TB230 was (2.21 ± 0.34) mol H 2 /mol glucose, which increased hydrogen production by over 40% compared with that of the wild type strain. The accumulation concentration of Fe (II) in the medium of the mutant strain TB230 with Fe (OH) 3 as the sole electron acceptor was (7.39 ± 0.49) mmol/l, which was approximately 3-fold greater than that of the wild type strain. The mutant strain TB230 showed high Fe (III)-reducing activity and hydrogen production by adopting glucose and pyruvate as the carbon source. In addition, the mutant strain TB230 was capable of Fe (III) reduction and hydrogen production under fresh or marine conditions. This result indicates that the mutant strain with high microbial Fe (III) reduction and hydrogen production is beneficial for the improvement of anaerobic performance. - Highlights: • The mutant strain TB230 was a transposon-mutagenized strain of Pantoea agglomerans BH18. • Strain TB230 was screened for high Fe (III)-reducing efficiency and hydrogen production. • H 2 yield and Fe (III)-reducing activity were 2.21 ± 0.34 and 7.39 ± 0.49 in marine condition. • Strain TB230 was capable of Fe (III) reduction and hydrogen production in fresh or marine condition

Phage transposon mutagenesis.

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Siegrist, M Sloan; Rubin, Eric J

2009-01-01

Phage transduction is an attractive method of genetic manipulation in mycobacteria. PhiMycoMarT7 is well suited for transposon mutagenesis as it is temperature sensitive for replication and contains T7 promoters that promote transcription, a highly active transposase gene, and an Escherichia coli oriR6 K origin of replication. Mycobacterial transposon mutant libraries produced by PhiMycoMarT7 transduction are amenable to both forward and reverse genetic studies. In this protocol, we detail the preparation of PhiMycoMarT7, including a description of the phage, reconstitution of the phage, purification of plaques, preparation of phage stock, and titering of phage stock. We then describe the transduction procedure and finally outline the isolation of individual transposon mutants.

Comprehensive identification of essential Staphylococcus aureus genes using Transposon-Mediated Differential Hybridisation (TMDH

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Burgis Timothy A

2009-07-01

Full Text Available Abstract Background In recent years there has been an increasing problem with Staphylococcus aureus strains that are resistant to treatment with existing antibiotics. An important starting point for the development of new antimicrobial drugs is the identification of "essential" genes that are important for bacterial survival and growth. Results We have developed a robust microarray and PCR-based method, Transposon-Mediated Differential Hybridisation (TMDH, that uses novel bioinformatics to identify transposon inserts in genome-wide libraries. Following a microarray-based screen, genes lacking transposon inserts are re-tested using a PCR and sequencing-based approach. We carried out a TMDH analysis of the S. aureus genome using a large random mariner transposon library of around a million mutants, and identified a total of 351 S. aureus genes important for survival and growth in culture. A comparison with the essential gene list experimentally derived for Bacillus subtilis highlighted interesting differences in both pathways and individual genes. Conclusion We have determined the first comprehensive list of S. aureus essential genes. This should act as a useful starting point for the identification of potential targets for novel antimicrobial compounds. The TMDH methodology we have developed is generic and could be applied to identify essential genes in other bacterial pathogens.

The Nucleoid Binding Protein H-NS Biases Genome-Wide Transposon Insertion Landscapes

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Satoshi Kimura

2016-08-01

Full Text Available Transposon insertion sequencing (TIS; also known as TnSeq is a potent approach commonly used to comprehensively define the genetic loci that contribute to bacterial fitness in diverse environments. A key presumption underlying analyses of TIS datasets is that loci with a low frequency of transposon insertions contribute to fitness. However, it is not known whether factors such as nucleoid binding proteins can alter the frequency of transposon insertion and thus whether TIS output may systematically reflect factors that are independent of the role of the loci in fitness. Here, we investigated whether the histone-like nucleoid structuring (H-NS protein, which preferentially associates with AT-rich sequences, modulates the frequency of Mariner transposon insertion in the Vibrio cholerae genome, using comparative analysis of TIS results from wild-type (wt and Δhns V. cholerae strains. These analyses were overlaid on gene classification based on GC content as well as on extant genome-wide identification of H-NS binding loci. Our analyses revealed a significant dearth of insertions within AT-rich loci in wt V. cholerae that was not apparent in the Δhns insertion library. Additionally, we observed a striking correlation between genetic loci that are overrepresented in the Δhns insertion library relative to their insertion frequency in wt V. cholerae and loci previously found to physically interact with H-NS. Collectively, our findings reveal that factors other than genetic fitness can systematically modulate the frequency of transposon insertions in TIS studies and add a cautionary note to interpretation of TIS data, particularly for AT-rich sequences.

Generation of mariner-based transposon insertion mutant library of Bacillus sphaericus 2297 and investigation of genes involved in sporulation and mosquito-larvicidal crystal protein synthesis.

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Wu, Yiming; Hu, Xiaomin; Ge, Yong; Zheng, Dasheng; Yuan, Zhiming

2012-05-01

Bacillus sphaericus has been used with great success in mosquito control programs worldwide. Under conditions of nutrient limitation, it undergoes sporulation via a series of well defined morphological stages. However, only a small number of genes involved in sporulation have been identified. To identify genes associated with sporulation, and to understand the relationship between sporulation and crystal protein synthesis, a random mariner-based transposon insertion mutant library of B. sphaericus strain 2297 was constructed and seven sporulation-defective mutants were selected. Sequencing of the DNA flanking of the transposon insertion identified several genes involved in sporulation. The morphologies of mutants were determined by electron microscopy and synthesis of crystal proteins was analyzed by SDS-PAGE and Western blot. Four mutants blocked at early stages of sporulation failed to produce crystal proteins and had lower larvicidal activity. However, the other three mutants were blocked at later stages and were able to form crystal proteins, and the larvicidal activity was similar to wild type. These results indicated that crystal protein synthesis in B. sphaericus is dependent on sporulation initiation. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Transposon Invasion of the Paramecium Germline Genome Countered by a Domesticated PiggyBac Transposase and the NHEJ Pathway

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Dubois, Emeline; Bischerour, Julien; Marmignon, Antoine; Mathy, Nathalie; Régnier, Vinciane; Bétermier, Mireille

2012-01-01

Sequences related to transposons constitute a large fraction of extant genomes, but insertions within coding sequences have generally not been tolerated during evolution. Thanks to their unique nuclear dimorphism and to their original mechanism of programmed DNA elimination from their somatic nucleus (macronucleus), ciliates are emerging model organisms for the study of the impact of transposable elements on genomes. The germline genome of the ciliate Paramecium, located in its micronucleus, contains thousands of short intervening sequences, the IESs, which interrupt 47% of genes. Recent data provided support to the hypothesis that an evolutionary link exists between Paramecium IESs and Tc1/mariner transposons. During development of the macronucleus, IESs are excised precisely thanks to the coordinated action of PiggyMac, a domesticated piggyBac transposase, and of the NHEJ double-strand break repair pathway. A PiggyMac homolog is also required for developmentally programmed DNA elimination in another ciliate, Tetrahymena. Here, we present an overview of the life cycle of these unicellular eukaryotes and of the developmentally programmed genome rearrangements that take place at each sexual cycle. We discuss how ancient domestication of a piggyBac transposase might have allowed Tc1/mariner elements to spread throughout the germline genome of Paramecium, without strong counterselection against insertion within genes. PMID:22888464

Random insertion and gene disruption via transposon mutagenesis of Ureaplasma parvum using a mini-transposon plasmid.

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Aboklaish, Ali F; Dordet-Frisoni, Emilie; Citti, Christine; Toleman, Mark A; Glass, John I; Spiller, O Brad

2014-11-01

While transposon mutagenesis has been successfully used for Mycoplasma spp. to disrupt and determine non-essential genes, previous attempts with Ureaplasma spp. have been unsuccessful. Using a polyethylene glycol-transformation enhancing protocol, we were able to transform three separate serovars of Ureaplasma parvum with a Tn4001-based mini-transposon plasmid containing a gentamicin resistance selection marker. Despite the large degree of homology between Ureaplasma parvum and Ureaplasma urealyticum, all attempts to transform the latter in parallel failed, with the exception of a single clinical U. urealyticum isolate. PCR probing and sequencing were used to confirm transposon insertion into the bacterial genome and identify disrupted genes. Transformation of prototype serovar 3 consistently resulted in transfer only of sequence between the mini-transposon inverted repeats, but some strains showed additional sequence transfer. Transposon insertion occurred randomly in the genome resulting in unique disruption of genes UU047, UU390, UU440, UU450, UU520, UU526, UU582 for single clones from a panel of screened clones. An intergenic insertion between genes UU187 and UU188 was also characterised. Two phenotypic alterations were observed in the mutated strains: Disruption of a DEAD-box RNA helicase (UU582) altered growth kinetics, while the U. urealyticum strain lost resistance to serum attack coincident with disruption of gene UUR10_137 and loss of expression of a 41 kDa protein. Transposon mutagenesis was used successfully to insert single copies of a mini-transposon into the genome and disrupt genes leading to phenotypic changes in Ureaplasma parvum strains. This method can now be used to deliver exogenous genes for expression and determine essential genes for Ureaplasma parvum replication in culture and experimental models. Copyright © 2014 Elsevier GmbH. All rights reserved.

Tc7, a Tc1-hitch hiking transposon in Caenorhabditis elegans.

OpenAIRE

Rezsohazy, R; van Luenen, H G; Durbin, R M; Plasterk, R H

1997-01-01

We have found a novel transposon in the genome of Caenorhabditis elegans. Tc7 is a 921 bp element, made up of two 345 bp inverted repeats separated by a unique, internal sequence. Tc7 does not contain an open reading frame. The outer 38 bp of the inverted repeat show 36 matches with the outer 38 bp of Tc1. This region of Tc1 contains the Tc1-transposase binding site. Furthermore, Tc7 is flanked by TA dinucleotides, just like Tc1, which presumably correspond to the target duplication generated...

[Active miniature inverted-repeat transposable elements transposon in plants: a review].

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Hu, Bingjie; Zhou, Mingbing

2018-02-25

Miniature inverted-repeat transposable elements transposon is a special transposon that could transpose by "cut-paste" mechanism, which is one of characteristics of DNA transposons. Otherwise, the copy number of MITEs is very high, which is one of characteristics of RNA transposons. Many MITE families have been reported, but little about active MITEs. We summarize recent advances in studying active MITEs. Most the MITEs belong to the Tourist-like family, such as mPing, mGing, PhTourist1, Tmi1 and PhTst-3. Additionally, DTstu1 and MITE-39 belong to Stowaway-like family, and AhMITEs1 belongs to Mutator-like family. Moreover, we summarize the structure (terminal inverse repeats and target site duplications), copy number, evolution pattern and transposition characteristics of these active MITEs, to provide the foundation for the identification of other active MITEs and subsequent research on MITE transposition and amplification mechanism.

Minos as a novel Tc1/mariner-type transposable element for functional genomic analysis in Aspergillus nidulans.

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Evangelinos, Minoas; Anagnostopoulos, Gerasimos; Karvela-Kalogeraki, Iliana; Stathopoulou, Panagiota M; Scazzocchio, Claudio; Diallinas, George

2015-08-01

Transposons constitute powerful genetic tools for gene inactivation, exon or promoter trapping and genome analyses. The Minos element from Drosophila hydei, a Tc1/mariner-like transposon, has proved as a very efficient tool for heterologous transposition in several metazoa. In filamentous fungi, only a handful of fungal-specific transposable elements have been exploited as genetic tools, with the impala Tc1/mariner element from Fusarium oxysporum being the most successful. Here, we developed a two-component transposition system to manipulate Minos transposition in Aspergillus nidulans (AnMinos). Our system allows direct selection of transposition events based on re-activation of niaD, a gene necessary for growth on nitrate as a nitrogen source. On average, among 10(8) conidiospores, we obtain up to ∼0.8×10(2) transposition events leading to the expected revertant phenotype (niaD(+)), while ∼16% of excision events lead to AnMinos loss. Characterized excision footprints consisted of the four terminal bases of the transposon flanked by the TA target duplication and led to no major DNA rearrangements. AnMinos transposition depends on the presence of its homologous transposase. Its frequency was not significantly affected by temperature, UV irradiation or the transcription status of the original integration locus (niaD). Importantly, transposition is dependent on nkuA, encoding an enzyme essential for non-homologous end joining of DNA in double-strand break repair. AnMinos proved to be an efficient tool for functional analysis as it seems to transpose in different genomic loci positions in all chromosomes, including a high proportion of integration events within or close to genes. We have used Minos to obtain morphological and toxic analogue resistant mutants. Interestingly, among morphological mutants some seem to be due to Minos-elicited over-expression of specific genes, rather than gene inactivation. Copyright © 2015 Elsevier Inc. All rights reserved.

TcA, the putative transposase of the C. elegans Tc1 transposon, has an N-terminal DNA binding domain.

OpenAIRE

Schukkink, R F; Plasterk, R H

1990-01-01

Tc1 is a transposon present in several copies in the genome of all natural isolates of the nematode C.elegans; it is actively transposing in many strains. In those strains Tc1 insertion is the main cause of spontaneous mutations. The transposon contains one large ORF that we call TcA; we assume that the TcA protein is the transposase of Tc1. We expressed TcA in E.coli, purified the protein and showed that it has a strong affinity for DNA (both single stranded and double stranded). A fusion pr...

Suicidal autointegration of sleeping beauty and piggyBac transposons in eukaryotic cells.

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Yongming Wang

2014-03-01

Full Text Available Transposons are discrete segments of DNA that have the distinctive ability to move and replicate within genomes across the tree of life. 'Cut and paste' DNA transposition involves excision from a donor locus and reintegration into a new locus in the genome. We studied molecular events following the excision steps of two eukaryotic DNA transposons, Sleeping Beauty (SB and piggyBac (PB that are widely used for genome manipulation in vertebrate species. SB originates from fish and PB from insects; thus, by introducing these transposons to human cells we aimed to monitor the process of establishing a transposon-host relationship in a naïve cellular environment. Similarly to retroviruses, neither SB nor PB is capable of self-avoidance because a significant portion of the excised transposons integrated back into its own genome in a suicidal process called autointegration. Barrier-to-autointegration factor (BANF1, a cellular co-factor of certain retroviruses, inhibited transposon autointegration, and was detected in higher-order protein complexes containing the SB transposase. Increasing size sensitized transposition for autointegration, consistent with elevated vulnerability of larger transposons. Both SB and PB were affected similarly by the size of the transposon in three different assays: excision, autointegration and productive transposition. Prior to reintegration, SB is completely separated from the donor molecule and followed an unbiased autointegration pattern, not associated with local hopping. Self-disruptive autointegration occurred at similar frequency for both transposons, while aberrant, pseudo-transposition events were more frequently observed for PB.

Remobilization of Sleeping Beauty transposons in the germline of Xenopus tropicalis

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Yergeau Donald A

2011-11-01

Full Text Available Abstract Background The Sleeping Beauty (SB transposon system has been used for germline transgenesis of the diploid frog, Xenopus tropicalis. Injecting one-cell embryos with plasmid DNA harboring an SB transposon substrate together with mRNA encoding the SB transposase enzyme resulted in non-canonical integration of small-order concatemers of the transposon. Here, we demonstrate that SB transposons stably integrated into the frog genome are effective substrates for remobilization. Results Transgenic frogs that express the SB10 transposase were bred with SB transposon-harboring animals to yield double-transgenic 'hopper' frogs. Remobilization events were observed in the progeny of the hopper frogs and were verified by Southern blot analysis and cloning of the novel integrations sites. Unlike the co-injection method used to generate founder lines, transgenic remobilization resulted in canonical transposition of the SB transposons. The remobilized SB transposons frequently integrated near the site of the donor locus; approximately 80% re-integrated with 3 Mb of the donor locus, a phenomenon known as 'local hopping'. Conclusions In this study, we demonstrate that SB transposons integrated into the X. tropicalis genome are effective substrates for excision and re-integration, and that the remobilized transposons are transmitted through the germline. This is an important step in the development of large-scale transposon-mediated gene- and enhancer-trap strategies in this highly tractable developmental model system.

Enzymatic engineering of the porcine genome with transposons and recombinases

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Carlson Daniel F

2007-07-01

Full Text Available Abstract Background Swine is an important agricultural commodity and biomedical model. Manipulation of the pig genome provides opportunity to improve production efficiency, enhance disease resistance, and add value to swine products. Genetic engineering can also expand the utility of pigs for modeling human disease, developing clinical treatment methodologies, or donating tissues for xenotransplantation. Realizing the full potential of pig genetic engineering requires translation of the complete repertoire of genetic tools currently employed in smaller model organisms to practical use in pigs. Results Application of transposon and recombinase technologies for manipulation of the swine genome requires characterization of their activity in pig cells. We tested four transposon systems- Sleeping Beauty, Tol2, piggyBac, and Passport in cultured porcine cells. Transposons increased the efficiency of DNA integration up to 28-fold above background and provided for precise delivery of 1 to 15 transgenes per cell. Both Cre and Flp recombinase were functional in pig cells as measured by their ability to remove a positive-negative selection cassette from 16 independent clones and over 20 independent genomic locations. We also demonstrated a Cre-dependent genetic switch capable of eliminating an intervening positive-negative selection cassette and activating GFP expression from episomal and genome-resident transposons. Conclusion We have demonstrated for the first time that transposons and recombinases are capable of mobilizing DNA into and out of the porcine genome in a precise and efficient manner. This study provides the basis for developing transposon and recombinase based tools for genetic engineering of the swine genome.

Identification of genes potentially involved in solute stress response in Sphingomonas wittichii RW1 by transposon mutant recovery

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Edith eCoronado

2014-11-01

Full Text Available The term water stress refers to the effects of low water availability on microbial growth and physiology. Water availability has been proposed as a major constraint for the use of microorganisms in contaminated sites with the purpose of bioremediation. Sphingomonas wittichii RW1 is a bacterium capable of degrading the xenobiotic compounds dibenzofuran and dibenzo-p-dioxin, and has potential to be used for targeted bioremediation. The aim of the current work was to identify genes implicated in water stress in RW1 by means of transposon mutagenesis and mutant growth experiments. Conditions of low water potential were mimicked by adding NaCl to the growth media. Three different mutant selection or separation method were tested, which, however recovered different mutants. Recovered transposon mutants with poorer growth under salt-induced water stress carried insertions in genes involved in proline and glutamate biosynthesis, and further in a gene putatively involved in aromatic compound catabolism. Transposon mutants growing poorer on medium with lowered water potential also included ones that had insertions in genes involved in more general functions such as transcriptional regulation, elongation factor, cell division protein, RNA polymerase β or an aconitase.

Genome-wide analysis of Tol2 transposon reintegration in zebrafish.

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Kondrychyn, Igor; Garcia-Lecea, Marta; Emelyanov, Alexander; Parinov, Sergey; Korzh, Vladimir

2009-09-08

Tol2, a member of the hAT family of transposons, has become a useful tool for genetic manipulation of model animals, but information about its interactions with vertebrate genomes is still limited. Furthermore, published reports on Tol2 have mainly been based on random integration of the transposon system after co-injection of a plasmid DNA harboring the transposon and a transposase mRNA. It is important to understand how Tol2 would behave upon activation after integration into the genome. We performed a large-scale enhancer trap (ET) screen and generated 338 insertions of the Tol2 transposon-based ET cassette into the zebrafish genome. These insertions were generated by remobilizing the transposon from two different donor sites in two transgenic lines. We found that 39% of Tol2 insertions occurred in transcription units, mostly into introns. Analysis of the transposon target sites revealed no strict specificity at the DNA sequence level. However, Tol2 was prone to target AT-rich regions with weak palindromic consensus sequences centered at the insertion site. Our systematic analysis of sequential remobilizations of the Tol2 transposon from two independent sites within a vertebrate genome has revealed properties such as a tendency to integrate into transcription units and into AT-rich palindrome-like sequences. This information will influence the development of various applications involving DNA transposons and Tol2 in particular.

Gene-specific cell labeling using MiMIC transposons.

Science.gov (United States)

Gnerer, Joshua P; Venken, Koen J T; Dierick, Herman A

2015-04-30

Binary expression systems such as GAL4/UAS, LexA/LexAop and QF/QUAS have greatly enhanced the power of Drosophila as a model organism by allowing spatio-temporal manipulation of gene function as well as cell and neural circuit function. Tissue-specific expression of these heterologous transcription factors relies on random transposon integration near enhancers or promoters that drive the binary transcription factor embedded in the transposon. Alternatively, gene-specific promoter elements are directly fused to the binary factor within the transposon followed by random or site-specific integration. However, such insertions do not consistently recapitulate endogenous expression. We used Minos-Mediated Integration Cassette (MiMIC) transposons to convert host loci into reliable gene-specific binary effectors. MiMIC transposons allow recombinase-mediated cassette exchange to modify the transposon content. We developed novel exchange cassettes to convert coding intronic MiMIC insertions into gene-specific binary factor protein-traps. In addition, we expanded the set of binary factor exchange cassettes available for non-coding intronic MiMIC insertions. We show that binary factor conversions of different insertions in the same locus have indistinguishable expression patterns, suggesting that they reliably reflect endogenous gene expression. We show the efficacy and broad applicability of these new tools by dissecting the cellular expression patterns of the Drosophila serotonin receptor gene family. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

A highly efficient transposon mutagenesis system for the tomato pathogen Clavibacter michiganensis subsp. michiganensis.

Science.gov (United States)

Kirchner, O; Gartemann, K H; Zellermann, E M; Eichenlaub, R; Burger, A

2001-11-01

A transposon mutagenesis system for Clavibacter michiganensis subsp. michiganensis was developed based on antibiotic resistance transposons that were derived from the insertion element IS1409 from Arthrobacter sp. strain TM1 NCIB12013. As a prerequisite, the electroporation efficiency was optimized by using unmethylated DNA and treatment of the cells with glycine such that about 5 x 10(6) transformants per microg of DNA were generally obtained. Electroporation of C. michiganensis subsp. michiganensis with a suicide vector carrying transposon Tn1409C resulted in approximately 1 x 10(3) transposon mutants per pg of DNA and thus is suitable for saturation mutagenesis. Analysis of Tn1409C insertion sites suggests a random mode of transposition. Transposition of Tn1409C was also demonstrated for other subspecies of C. michiganensis.

Genome-wide analysis of Tol2 transposon reintegration in zebrafish

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Parinov Sergey

2009-09-01

Full Text Available Abstract Background Tol2, a member of the hAT family of transposons, has become a useful tool for genetic manipulation of model animals, but information about its interactions with vertebrate genomes is still limited. Furthermore, published reports on Tol2 have mainly been based on random integration of the transposon system after co-injection of a plasmid DNA harboring the transposon and a transposase mRNA. It is important to understand how Tol2 would behave upon activation after integration into the genome. Results We performed a large-scale enhancer trap (ET screen and generated 338 insertions of the Tol2 transposon-based ET cassette into the zebrafish genome. These insertions were generated by remobilizing the transposon from two different donor sites in two transgenic lines. We found that 39% of Tol2 insertions occurred in transcription units, mostly into introns. Analysis of the transposon target sites revealed no strict specificity at the DNA sequence level. However, Tol2 was prone to target AT-rich regions with weak palindromic consensus sequences centered at the insertion site. Conclusion Our systematic analysis of sequential remobilizations of the Tol2 transposon from two independent sites within a vertebrate genome has revealed properties such as a tendency to integrate into transcription units and into AT-rich palindrome-like sequences. This information will influence the development of various applications involving DNA transposons and Tol2 in particular.

IS30-related transposon mediated insertional inactivation of bile salt hydrolase (bsh1) gene of Lactobacillus plantarum strain Lp20.

Science.gov (United States)

Kumar, Rajesh; Grover, Sunita; Kaushik, Jai K; Batish, Virender Kumar

2014-01-01

Lactobacillus plantarum is a flexible and versatile microorganism that inhabits a variety of niches, and its genome may express up to four bsh genes to maximize its survival in the mammalian gut. However, the ecological significance of multiple bsh genes in L. plantarum is still not clearly understood. Hence, this study demonstrated the disruption of bile salt hydrolase (bsh1) gene due to the insertion of a transposable element in L. plantarum Lp20 - a wild strain of human fecal origin. Surprisingly, L. plantarum strain Lp20 produced a ∼2.0 kb bsh1 amplicon against the normal size (∼1.0 kb) bsh1 amplicon of Bsh(+)L. plantarum Lp21. Strain Lp20 exhibited minimal Bsh activity in spite of having intact bsh2, bsh3 and bsh4 genes in its genome and hence had a Bsh(-) phenotype. Cloning and sequence characterization of Lp20 bsh1 gene predicted four individual open reading frames (ORFs) within this region. BLAST analysis of ORF1 and ORF2 revealed significant sequence similarity to the L. plantarum bsh1 gene while ORF3 and ORF4 showed high sequence homology to IS30-family transposases. Since, IS30-related transposon element was inserted within Lp20 bsh1 gene in reverse orientation (3'-5'), it introduced several stop codons and disrupted the protein reading frames of both Bsh1 and transposase. Inverted terminal repeats (GGCAGATTG) of transposon, mediated its insertion at 255-263 nt and 1301-1309 nt positions of Lp20 bsh1 gene. In conclusion, insertion of IS30 related-transposon within the bsh1 gene sequence of L. plantarum strain Lp20 demolished the integrity and functionality of Bsh1 enzyme. Additionally, this transposon DNA sequence remains active among various Lactobacillus spp. and hence harbors the potential to be explored in the development of efficient insertion mutagenesis system. Copyright © 2013 Elsevier GmbH. All rights reserved.

A putative autonomous 20.5 kb-CACTA transposon insertion in an F3'H allele identifies a new CACTA transposon subfamily in Glycine max

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Vodkin Lila

2008-12-01

Full Text Available Abstract Background The molecular organization of very few genetically defined CACTA transposon systems have been characterized thoroughly as those of Spm/En in maize, Tam1 of Antirrhinum majus Candystripe1 (Cs1 from Sorghum bicolor and CAC1 from Arabidopsis thaliana, for example. To date, only defective deletion derivatives of CACTA elements have been described for soybean, an economically important plant species whose genome sequence will be completed in 2008. Results We identified a 20.5 kb insertion in a soybean flavonoid 3'-hydroxylase (F3'H gene representing the t* allele (stable gray trichome color whose origin traces to a single mutable chimeric plant displaying both tawny and gray trichomes. This 20.5 kb insertion has the molecular structure of a putative autonomous transposon of the CACTA family, designated Tgmt*. It encodes a large gene that was expressed in two sister isolines (T* and tm of the stable gray line (t* from which Tgmt* was isolated. RT-PCR derived cDNAs uncovered the structure of a large precursor mRNA as well as alternatively spliced transcripts reminiscent of the TNPA-mRNA generated by the En-1 element of maize but without sequence similarity to the maize TNPA. The larger mRNA encodes a transposase with a tnp2 and TNP1-transposase family domains. Because the two soybean lines expressing Tgmt* were derived from the same mutable chimeric plant that created the stable gray trichome t* allele line from which the element was isolated, Tgmt* has the potential to be an autonomous element that was rapidly inactivated in the stable gray trichome t* line. Comparison of Tgmt* to previously described Tgm elements demonstrated that two subtypes of CACTA transposon families exist in soybean based on divergence of their characteristic subterminal repeated motifs and their transposases. In addition, we report the sequence and annotation of a BAC clone containing the F3'H gene (T locus which was interrupted by the novel Tgmt* element

Identifying transposon insertions and their effects from RNA-sequencing data.

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de Ruiter, Julian R; Kas, Sjors M; Schut, Eva; Adams, David J; Koudijs, Marco J; Wessels, Lodewyk F A; Jonkers, Jos

2017-07-07

Insertional mutagenesis using engineered transposons is a potent forward genetic screening technique used to identify cancer genes in mouse model systems. In the analysis of these screens, transposon insertion sites are typically identified by targeted DNA-sequencing and subsequently assigned to predicted target genes using heuristics. As such, these approaches provide no direct evidence that insertions actually affect their predicted targets or how transcripts of these genes are affected. To address this, we developed IM-Fusion, an approach that identifies insertion sites from gene-transposon fusions in standard single- and paired-end RNA-sequencing data. We demonstrate IM-Fusion on two separate transposon screens of 123 mammary tumors and 20 B-cell acute lymphoblastic leukemias, respectively. We show that IM-Fusion accurately identifies transposon insertions and their true target genes. Furthermore, by combining the identified insertion sites with expression quantification, we show that we can determine the effect of a transposon insertion on its target gene(s) and prioritize insertions that have a significant effect on expression. We expect that IM-Fusion will significantly enhance the accuracy of cancer gene discovery in forward genetic screens and provide initial insight into the biological effects of insertions on candidate cancer genes. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

Transposon domestication versus mutualism in ciliate genome rearrangements.

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Alexander Vogt

Full Text Available Ciliated protists rearrange their genomes dramatically during nuclear development via chromosome fragmentation and DNA deletion to produce a trimmer and highly reorganized somatic genome. The deleted portion of the genome includes potentially active transposons or transposon-like sequences that reside in the germline. Three independent studies recently showed that transposase proteins of the DDE/DDD superfamily are indispensible for DNA processing in three distantly related ciliates. In the spirotrich Oxytricha trifallax, high copy-number germline-limited transposons mediate their own excision from the somatic genome but also contribute to programmed genome rearrangement through a remarkable transposon mutualism with the host. By contrast, the genomes of two oligohymenophorean ciliates, Tetrahymena thermophila and Paramecium tetraurelia, encode homologous PiggyBac-like transposases as single-copy genes in both their germline and somatic genomes. These domesticated transposases are essential for deletion of thousands of different internal sequences in these species. This review contrasts the events underlying somatic genome reduction in three different ciliates and considers their evolutionary origins and the relationships among their distinct mechanisms for genome remodeling.

Identification and evolutionary dynamics of cacta DNA transposons in brassica

International Nuclear Information System (INIS)

Nouroz, F.; Noreen, S.; Harrison, J.S.H.

2017-01-01

Transposable elements are the major drivers of genome evolution and plasticity. Due to their transposition mode, they are classified into two major classes as Retrotransposons and DNA transposons. The En/Spm or CACTA elements are diverse group of DNA transposons proliferating in plant genomes. Various bioinformatics and molecular approaches were used for identification and distribution of CACTA transposons in Brassica genome. A combination of dot plot analysis and BLASTN searches yielded 35 autonomous and 7 non-autonomous CACTA elements in Brassica. The elements ranged in sizes from 1.2 kb non-autonomous elements to 11kb autonomous elements, terminated by 3 bp Target Site Duplication (TSD) and ~15 bp conserved Terminal Inverted Repeat (TIR) motifs (5'-CACTACAAGAAAACA-3'), with heterogeneous internal regions. The transposase (TNP) was identified from autonomous CACTA elements, while other protein domains from Brassica and other plants CACTA revealed similar organizations with minor differences. Both transposases (TNPD, TNPA) are present in most CACTA, while a few CACTA harboured an additional ATHILA ORF1-like domain. The PCR analysis amplified the CACTA transposases from 40 Brassica accessions (A, B, and C-genome) suggesting their distribution among various Brassica crops. A detailed characterization and evolutionary analysis of the identified CACTA elements allowed some to be placed in genome-specific groups, while most of them (Brassica-Arabidopsis elements) have followed the same evolutionary line. The distribution of CACTA in Brassica concluded that 3 bp TSDs generating CACTA transposons contributed significantly to genome size and evolution of Brassica genome. (author)

Analysis of an ordered, comprehensive STM mutant library in infectious Borrelia burgdorferi: insights into the genes required for mouse infectivity.

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Tao Lin

Full Text Available The identification of genes important in the pathogenesis of Lyme disease Borrelia has been hampered by exceedingly low transformation rates in low-passage, infectious organisms. Using the infectious, moderately transformable B. burgdorferi derivative 5A18NP1 and signature-tagged versions of the Himar1 transposon vector pGKT, we have constructed a defined transposon library for the efficient genome-wide investigation of genes required for wild-type pathogenesis, in vitro growth, physiology, morphology, and plasmid replication. To facilitate analysis, the insertion sites of 4,479 transposon mutants were determined by sequencing. The transposon insertions were widely distributed across the entire B. burgdorferi genome, with an average of 2.68 unique insertion sites per kb DNA. The 10 linear plasmids and 9 circular plasmids had insertions in 33 to 100 percent of their predicted genes. In contrast, only 35% of genes in the 910 kb linear chromosome had incapacitating insertions; therefore, the remaining 601 chromosomal genes may represent essential gene candidates. In initial signature-tagged mutagenesis (STM analyses, 434 mutants were examined at multiple tissue sites for infectivity in mice using a semi-quantitative, Luminex-based DNA detection method. Examples of genes found to be important in mouse infectivity included those involved in motility, chemotaxis, the phosphoenolpyruvate phosphotransferase system, and other transporters, as well as putative plasmid maintenance genes. Availability of this ordered STM library and a high-throughput screening method is expected to lead to efficient assessment of the roles of B. burgdorferi genes in the infectious cycle and pathogenesis of Lyme disease.

Application of In Vitro Transposon Mutagenesis to Erythromycin Strain Improvement in Saccharopolyspora erythraea.

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Weber, J Mark; Reeves, Andrew; Cernota, William H; Wesley, Roy K

2017-01-01

Transposon mutagenesis is an invaluable technique in molecular biology for the creation of random mutations that can be easily identified and mapped. However, in the field of microbial strain improvement, transposon mutagenesis has scarcely been used; instead, chemical and physical mutagenic methods have been traditionally favored. Transposons have the advantage of creating single mutations in the genome, making phenotype to genotype assignments less challenging than with traditional mutagens which commonly create multiple mutations in the genome. The site of a transposon mutation can also be readily mapped using DNA sequencing primer sites engineered into the transposon termini. In this chapter an in vitro method for transposon mutagenesis of Saccharopolyspora erythraea is presented. Since in vivo transposon tools are not available for most actinomycetes including S. erythraea, an in vitro method was developed. The in vitro method involves a significant investment in time and effort to create the mutants, but once the mutants are made and screened, a large number of highly relevant mutations of direct interest to erythromycin production can be found.

A plasmid-transposon hybrid mutagenesis system effective in a broad range of Enterobacteria

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Rita eMonson

2015-12-01

Full Text Available Random transposon mutagenesis is a powerful technique used to generate libraries of genetic insertions in many different bacterial strains. Here we develop a system facilitating random transposon mutagenesis in a range of different Gram-negative bacterial strains, including Pectobacterium atrosepticum, Citrobacter rodentium, Serratia sp. ATCC39006, Serratia plymuthica, Dickeya dadantii and many more. Transposon mutagenesis was optimized in each of these strains and three studies are presented to show the efficacy of this system. Firstly, the important agricultural pathogen D. dadantii was mutagenized. Two mutants that showed reduced protease production and one mutant producing the previously cryptic pigment, indigoidine, were identified and characterized. Secondly, the enterobacterium, Serratia sp. ATCC39006 was mutagenized and mutants incapable of producing gas vesicles, proteinaceous intracellular organelles, were identified. One of these contained a β-galactosidase transcriptional fusion within the gene gvpA1, essential for gas vesicle production. Finally, the system was used to mutate the biosynthetic gene clusters of the antifungal, anti-oomycete and anticancer polyketide, oocydin A, in the plant-associated enterobacterium, Dickeya solani MK10. The mutagenesis system was developed to allow easy identification of transposon insertion sites by sequencing, after facile generation of a replicon encompassing the transposon and adjacent DNA, post-excision. Furthermore, the system can also create transcriptional fusions with either β-galactosidase or β-glucuronidase as reporters, and exploits a variety of drug resistance markers so that multiple selectable fusions can be generated in a single strain. This system of various transposons has wide utility and can be combined in many different ways.

Transposons As Tools for Functional Genomics in Vertebrate Models.

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Kawakami, Koichi; Largaespada, David A; Ivics, Zoltán

2017-11-01

Genetic tools and mutagenesis strategies based on transposable elements are currently under development with a vision to link primary DNA sequence information to gene functions in vertebrate models. By virtue of their inherent capacity to insert into DNA, transposons can be developed into powerful tools for chromosomal manipulations. Transposon-based forward mutagenesis screens have numerous advantages including high throughput, easy identification of mutated alleles, and providing insight into genetic networks and pathways based on phenotypes. For example, the Sleeping Beauty transposon has become highly instrumental to induce tumors in experimental animals in a tissue-specific manner with the aim of uncovering the genetic basis of diverse cancers. Here, we describe a battery of mutagenic cassettes that can be applied in conjunction with transposon vectors to mutagenize genes, and highlight versatile experimental strategies for the generation of engineered chromosomes for loss-of-function as well as gain-of-function mutagenesis for functional gene annotation in vertebrate models, including zebrafish, mice, and rats. Copyright © 2017 Elsevier Ltd. All rights reserved.

Bursts and horizontal evolution of DNA transposons in the speciation of pseudotetraploid salmonids

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Davidson William S

2007-11-01

Full Text Available Abstract Background Several genome duplications have occurred in the evolutionary history of teleost fish. In returning to a stable diploid state, the polyploid genome reorganized, and large portions are lost, while the fish lines evolved to numerous species. Large scale transposon movement has been postulated to play an important role in the genome reorganization process. We analyzed the DNA sequence of several large loci in Salmo salar and other species for the presence of DNA transposon families. Results We have identified bursts of activity of 14 families of DNA transposons (12 Tc1-like and 2 piggyBac-like families, including 11 novel ones in genome sequences of Salmo salar. Several of these families have similar sequences in a number of closely and distantly related fish, lamprey, and frog species as well as in the parasite Schistosoma japonicum. Analysis of sequence similarities between copies within the families of these bursts demonstrates several waves of transposition activities coinciding with salmonid species divergence. Tc1-like families show a master gene-like copying process, illustrated by extensive but short burst of copying activity, while the piggyBac-like families show a more random copying pattern. Recent families may include copies with an open reading frame for an active transposase enzyme. Conclusion We have identified defined bursts of transposon activity that make use of master-slave and random mechanisms. The bursts occur well after hypothesized polyploidy events and coincide with speciation events. Parasite-mediated lateral transfer of transposons are implicated.

A comparative phylogenetic analysis of full-length mariner elements

Indian Academy of Sciences (India)

Mariner like elements (MLEs) are widely distributed type II transposons with an open reading frame (ORF) for transposase. We studied comparative phylogenetic evolution and inverted terminal repeat (ITR) conservation of MLEs from Indian saturniid silkmoth, Antheraea mylitta with other full length MLEs submitted in the ...

Stochastic Predator-Prey Dynamics of Transposons in the Human Genome

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Xue, Chi; Goldenfeld, Nigel

2016-11-01

Transposable elements, or transposons, are DNA sequences that can jump from site to site in the genome during the life cycle of a cell, usually encoding the very enzymes which perform their excision. However, some transposons are parasitic, relying on the enzymes produced by the regular transposons. In this case, we show that a stochastic model, which takes into account the small copy numbers of the active transposons in a cell, predicts noise-induced predator-prey oscillations with a characteristic time scale that is much longer than the cell replication time, indicating that the state of the predator-prey oscillator is stored in the genome and transmitted to successive generations. Our work demonstrates the important role of the number fluctuations in the expression of mobile genetic elements, and shows explicitly how ecological concepts can be applied to the dynamics and fluctuations of living genomes.

An Enterobacter plasmid as a new genetic background for the transposon Tn1331

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Alavi MR

2011-11-01

Full Text Available Mohammad R Alavi1,2, Vlado Antonic2, Adrien Ravizee1, Peter J Weina3, Mina Izadjoo1,2, Alexander Stojadinovic21Division of Wound Biology and Translational Research, Armed Forces Institute of Pathology and American Registry of Pathology, Washington DC, 2Combat Wound Initiative Program, Walter Reed Army Medical Center, Washington DC, 3The Walter Reed Army Institute of Research, Silver Spring, MD, USABackground: Genus Enterobacter includes important opportunistic nosocomial pathogens that could infect complex wounds. The presence of antibiotic resistance genes in these microorganisms represents a challenging clinical problem in the treatment of these wounds. In the authors’ screening of antibiotic-resistant bacteria from complex wounds, an Enterobacter species was isolated that harbors antibiotic-resistant plasmids conferring resistance to Escherichia coli. The aim of this study was to identify the resistance genes carried by one of these plasmids.Methods: The plasmids from the Enterobacter isolate were propagated in E. coli and one of the plasmids, designated as pR23, was sequenced by the Sanger method using fluorescent dye-terminator chemistry on a genetic analyzer. The assembled sequence was annotated by search of the GenBank database.Results: Plasmid pR23 is composed of the transposon Tn1331 and a backbone plasmid that is identical to the plasmid pPIGDM1 from Enterobacter agglomerans. The multidrug-resistance transposon Tn1331, which confers resistance to aminoglycoside and beta lactam antibiotics, has been previously isolated only from Klebsiella. The Enterobacter plasmid pPIGDM1, which carries a ColE1-like origin of replication and has no apparent selective marker, appears to provide a backbone for propagation of Tn1331 in Enterobacter. The recognition sequence of Tn1331 transposase for insertion into pPIGDM1 is the pentanucleotide TATTA, which occurs only once throughout the length of this plasmid.Conclusion: Transposition of Tn1331 into

Construction of a large-scale Burkholderia cenocepacia J2315 transposon mutant library

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Wong, Yee-Chin; Pain, Arnab; Nathan, Sheila

2014-09-01

Burkholderia cenocepacia, a pathogenic member of the Burkholderia cepacia complex (Bcc), has emerged as a significant threat towards cystic fibrosis patients, where infection often leads to the fatal clinical manifestation known as cepacia syndrome. Many studies have investigated the pathogenicity of B. cenocepacia as well as its ability to become highly resistant towards many of the antibiotics currently in use. In addition, studies have also been undertaken to understand the pathogen's capacity to adapt and survive in a broad range of environments. Transposon based mutagenesis has been widely used in creating insertional knock-out mutants and coupled with recent advances in sequencing technology, robust tools to study gene function in a genome-wide manner have been developed based on the assembly of saturated transposon mutant libraries. In this study, we describe the construction of a large-scale library of B. cenocepacia transposon mutants. To create transposon mutants of B. cenocepacia strain J2315, electrocompetent bacteria were electrotransformed with the EZ-Tn5 transposome. Tetracyline resistant colonies were harvested off selective agar and pooled. Mutants were generated in multiple batches with each batch consisting of ˜20,000 to 40,000 mutants. Transposon insertion was validated by PCR amplification of the transposon region. In conclusion, a saturated B. cenocepacia J2315 transposon mutant library with an estimated total number of 500,000 mutants was successfully constructed. This mutant library can now be further exploited as a genetic tool to assess the function of every gene in the genome, facilitating the discovery of genes important for bacterial survival and adaptation, as well as virulence.

No-Go Zones for Mariner Transposition

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Brian J. Akerley

2016-10-01

Full Text Available The property of transposons to randomly insert into target DNA has long been exploited for generalized mutagenesis and forward genetic screens. Newer applications that monitor the relative abundance of each transposon insertion in large libraries of mutants can be used to evaluate the roles in cellular fitness of all genes of an organism, provided that transposition is in fact random across all genes. In a recent article, Kimura and colleagues identified an important exception to the latter assumption [S. Kimura, T. P. Hubbard, B. M. Davis, M. K. Waldor, mBio 7(4:e01351-16, 2016, doi:10.1128/mBio.01351-16]. They provide evidence that the Mariner transposon exhibits locus-specific site preferences in the presence of the histone-like nucleoid structuring protein H-NS. This effect was shown to bias results for important virulence loci in Vibrio cholerae and to result in misidentification of genes involved in growth in vitro. Fortunately, the bulk of this bacterium’s genome was unaffected by this bias, and recognizing the H-NS effect allows filtering to improve the accuracy of the results.

Evaluating Risks of Insertional Mutagenesis by DNA Transposons in Gene Therapy

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Hackett, Perry B.; Largaespada, David A.; Switzer, Kirsten C.; Cooper, Laurence J.N.

2013-01-01

Investigational therapy can be successfully undertaken using viral- and non-viral-mediated ex vivo gene transfer. Indeed, recent clinical trials have established the potential for genetically modified T cells to improve and restore health. Recently the Sleeping Beauty (SB) transposon/transposase system has been applied in clinical trials to stably insert a chimeric antigen receptor (CAR) to redirect T-cell specificity. We discuss the context in which the SB system can be harnessed for gene therapy and describe the human application of SB-modified CAR+ T cells. We have focused on theoretical issues relating to insertional mutagenesis in the context of human genomes that are naturally subjected to remobilization of transposons and the experimental evidence over the last decade of employing SB transposons for defining genes that induce cancer. These findings are put into the context of the use of SB transposons in the treatment of human disease. PMID:23313630

Transposon mutagenesis in Bifidobacterium breve: construction and characterization of a Tn5 transposon mutant library for Bifidobacterium breve UCC2003.

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Ruiz, Lorena; Motherway, Mary O'Connell; Lanigan, Noreen; van Sinderen, Douwe

2013-01-01

Bifidobacteria are claimed to contribute positively to human health through a range of beneficial or probiotic activities, including amelioration of gastrointestinal and metabolic disorders, and therefore this particular group of gastrointestinal commensals has enjoyed increasing industrial and scientific attention in recent years. However, the molecular mechanisms underlying these probiotic mechanisms are still largely unknown, mainly due to the fact that molecular tools for bifidobacteria are rather poorly developed, with many strains lacking genetic accessibility. In this work, we describe the generation of transposon insertion mutants in two bifidobacterial strains, B. breve UCC2003 and B. breve NCFB2258. We also report the creation of the first transposon mutant library in a bifidobacterial strain, employing B. breve UCC2003 and a Tn5-based transposome strategy. The library was found to be composed of clones containing single transposon insertions which appear to be randomly distributed along the genome. The usefulness of the library to perform phenotypic screenings was confirmed through identification and analysis of mutants defective in D-galactose, D-lactose or pullulan utilization abilities.

Transposon mutagenesis in Bifidobacterium breve: construction and characterization of a Tn5 transposon mutant library for Bifidobacterium breve UCC2003.

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Lorena Ruiz

Full Text Available Bifidobacteria are claimed to contribute positively to human health through a range of beneficial or probiotic activities, including amelioration of gastrointestinal and metabolic disorders, and therefore this particular group of gastrointestinal commensals has enjoyed increasing industrial and scientific attention in recent years. However, the molecular mechanisms underlying these probiotic mechanisms are still largely unknown, mainly due to the fact that molecular tools for bifidobacteria are rather poorly developed, with many strains lacking genetic accessibility. In this work, we describe the generation of transposon insertion mutants in two bifidobacterial strains, B. breve UCC2003 and B. breve NCFB2258. We also report the creation of the first transposon mutant library in a bifidobacterial strain, employing B. breve UCC2003 and a Tn5-based transposome strategy. The library was found to be composed of clones containing single transposon insertions which appear to be randomly distributed along the genome. The usefulness of the library to perform phenotypic screenings was confirmed through identification and analysis of mutants defective in D-galactose, D-lactose or pullulan utilization abilities.

A systematic identification of Kolobok superfamily transposons in Trichomonas vaginalis and sequence analysis on related transposases

Institute of Scientific and Technical Information of China (English)

Qingshu Meng; Kaifu Chen; Lina Ma; Songnian Hu; Jun Yu

2011-01-01

Transposons are sequence elements widely distributed among genomes of all three kingdoms of life, providing genomic changes and playing significant roles in genome evolution. Trichomonas vaginalis is an excellent model system for transposon study since its genome ( ～ 160 Mb) has been sequenced and is composed of ～65% transposons and other repetitive elements. In this study, we primarily report the identification of Kolobok-type transposons (termed tvBac) in T. vaginalis and the results of transposase sequence analysis. We categorized 24 novel subfamilies of the Kolobok element, including one autonomous subfamily and 23 non-autonomous subfamilies. We also identified a novel H2CH motif in tvBac transposases based on multiple sequence alignment. In addition, we supposed that tvBac and Mutator transposons may have evolved independently from a common ancestor according to our phylogenetic analysis. Our results provide basic information for the understanding of the function and evolution of tvBac transposons in particular and other related transposon families in general.

An inducible tool for random mutagenesis in Aspergillus niger based on the transposon Vader.

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Paun, Linda; Nitsche, Benjamin; Homan, Tim; Ram, Arthur F; Kempken, Frank

2016-07-01

The ascomycete Aspergillus niger is widely used in the biotechnology, for instance in producing most of the world's citric acid. It is also known as a major food and feed contaminant. While generation of gene knockouts for functional genomics has become feasible in ku70 mutants, analyzing gene functions or metabolic pathways remains a laborious task. An unbiased transposon-based mutagenesis approach may aid this process of analyzing gene functions by providing mutant libraries in a short time. The Vader transposon is a non-autonomous DNA-transposon, which is activated by the homologous tan1-transposase. However, in the most commonly used lab strain of A. niger (N400 strain and derivatives), we found that the transposase, encoded by the tan1 gene, is mutated and inactive. To establish a Vader transposon-based mutagenesis system in the N400 background, we expressed the functional transposase of A. niger strain CBS 513.88 under the control of an inducible promoter based on the Tet-on system, which is activated in the presence of the antibiotic doxycycline (DOX). Increasing amounts of doxycycline lead to higher Vader excision frequencies, whereas little to none activity of Vader was observed without addition of doxycycline. Hence, this system appears to be suitable for producing stable mutants in the A. niger N400 background.

Epigenetics and Evolution: Transposons and the Stochastic Epigenetic Modification Model

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Sergio Branciamore

2015-04-01

Full Text Available In addition to genetic variation, epigenetic variation and transposons can greatly affect the evolutionary fitnesses landscape and gene expression. Previously we proposed a mathematical treatment of a general epigenetic variation model that we called Stochastic Epigenetic Modification (SEM model. In this study we follow up with a special case, the Transposon Silencing Model (TSM, with, once again, emphasis on quantitative treatment. We have investigated the evolutionary effects of epigenetic changes due to transposon (T insertions; in particular, we have considered a typical gene locus A and postulated that (i the expression level of gene A depends on the epigenetic state (active or inactive of a cis- located transposon element T, (ii stochastic variability in the epigenetic silencing of T occurs only in a short window of opportunity during development, (iii the epigenetic state is then stable during further development, and (iv the epigenetic memory is fully reset at each generation. We develop the model using two complementary approaches: a standard analytical population genetics framework (di usion equations and Monte-Carlo simulations. Both approaches led to similar estimates for the probability of fixation and time of fixation of locus TA with initial frequency P in a randomly mating diploid population of effective size Ne. We have ascertained the e ect that ρ, the probability of transposon Modification during the developmental window, has on the population (species. One of our principal conclusions is that as ρ increases, the pattern of fixation of the combined TA locus goes from "neutral" to "dominant" to "over-dominant". We observe that, under realistic values of ρ, epigenetic Modifications can provide an e cient mechanism for more rapid fixation of transposons and cis-located gene alleles. The results obtained suggest that epigenetic silencing, even if strictly transient (being reset at each generation, can still have signi cant

Identification of novel genes responsible for ethanol and/or thermotolerance by transposon mutagenesis in Saccharomyces cerevisiae

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Kim, Hyun-Soo [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Sciences; Kim, Na-Rae [Ewha Womans Univ., Seoul (Korea, Republic of). Div. of Life and Pharmaceutical Sciences; Yang, Jungwoo [Ewha Womans Univ., Seoul (Korea, Republic of). Microbial Resources Research Center; Choi, Wonja [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Sciences; Ewha Womans Univ., Seoul (Korea, Republic of). Div. of Life and Pharmaceutical Sciences; Ewha Womans Univ., Seoul (Korea, Republic of). Microbial Resources Research Center

2011-08-15

Saccharomyces cerevisiae strains tolerant to ethanol and heat stresses are important for industrial ethanol production. In this study, five strains (Tn 1-5) tolerant to up to 15% ethanol were isolated by screening a transposon-mediated mutant library. Two of them displayed tolerance to heat (42 C). The determination of transposon insertion sites and Northern blot analysis identified seven putative genes (CMP2, IMD4, SSK2, PPG1, DLD3, PAM1, and MSN2) and revealed simultaneous down-regulations of CMP2 and IMD4, and SSK2 and PPG1, down-regulation of DLD3, and disruptions of the open reading frame of PAM1 and MSN2, indicating that ethanol and/or heat tolerance can be conferred. Knockout mutants of these seven individual genes were ethanol tolerant and three of them (SSK2, PPG1, and PAM1) were tolerant to heat. Such tolerant phenotypes reverted to sensitive phenotypes by the autologous or overexpression of each gene. Five transposon mutants showed higher ethanol production and grew faster than the control strain when cultured in rich media containing 30% glucose and initial 6% ethanol at 30 C. Of those, two thermotolerant transposon mutants (Tn 2 and Tn 3) exhibited significantly enhanced growth and ethanol production compared to the control at 42 C. The genes identified in this study may provide a basis for the application in developing industrial yeast strains. (orig.)

Hybrid Lentivirus-transposon Vectors With a Random Integration Profile in Human Cells

DEFF Research Database (Denmark)

Staunstrup, Nicklas H; Moldt, Brian; Mátés, Lajos

2009-01-01

Gene delivery by human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors (LVs) is efficient, but genomic integration of the viral DNA is strongly biased toward transcriptionally active loci resulting in an increased risk of insertional mutagenesis in gene therapy protocols. Nonviral...... Sleeping Beauty (SB) transposon vectors have a significantly safer insertion profile, but efficient delivery into relevant cell/tissue types is a limitation. In an attempt to combine the favorable features of the two vector systems we established a novel hybrid vector technology based on SB transposase......-mediated insertion of lentiviral DNA circles generated during transduction of target cells with integrase (IN)-defective LVs (IDLVs). By construction of a lentivirus-transposon hybrid vector allowing transposition exclusively from circular viral DNA substrates, we demonstrate that SB transposase added in trans...

Sequence-indexed mutations in maize using the UniformMu transposon-tagging population

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Baier John

2007-05-01

Full Text Available Abstract Background Gene knockouts are a critical resource for functional genomics. In Arabidopsis, comprehensive knockout collections were generated by amplifying and sequencing genomic DNA flanking insertion mutants. These Flanking Sequence Tags (FSTs map each mutant to a specific locus within the genome. In maize, FSTs have been generated using DNA transposons. Transposable elements can generate unstable insertions that are difficult to analyze for simple knockout phenotypes. Transposons can also generate somatic insertions that fail to segregate in subsequent generations. Results Transposon insertion sites from 106 UniformMu FSTs were tested for inheritance by locus-specific PCR. We confirmed 89% of the FSTs to be germinal transposon insertions. We found no evidence for somatic insertions within the 11% of insertion sites that were not confirmed. Instead, this subset of insertion sites had errors in locus-specific primer design due to incomplete or low-quality genomic sequences. The locus-specific PCR assays identified a knockout of a 6-phosphogluconate dehydrogenase gene that co-segregates with a seed mutant phenotype. The mutant phenotype linked to this knockout generates novel hypotheses about the role for the plastid-localized oxidative pentose phosphate pathway during grain-fill. Conclusion We show that FSTs from the UniformMu population identify stable, germinal insertion sites in maize. Moreover, we show that these sequence-indexed mutations can be readily used for reverse genetic analysis. We conclude from these data that the current collection of 1,882 non-redundant insertion sites from UniformMu provide a genome-wide resource for reverse genetics.

The expanding universe of transposon technologies for gene and cell engineering

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Ivics Zoltán

2010-12-01

Full Text Available Abstract Transposable elements can be viewed as natural DNA transfer vehicles that, similar to integrating viruses, are capable of efficient genomic insertion. The mobility of class II transposable elements (DNA transposons can be controlled by conditionally providing the transposase component of the transposition reaction. Thus, a DNA of interest (be it a fluorescent marker, a small hairpin (shRNA expression cassette, a mutagenic gene trap or a therapeutic gene construct cloned between the inverted repeat sequences of a transposon-based vector can be used for stable genomic insertion in a regulated and highly efficient manner. This methodological paradigm opened up a number of avenues for genome manipulations in vertebrates, including transgenesis for the generation of transgenic cells in tissue culture, the production of germline transgenic animals for basic and applied research, forward genetic screens for functional gene annotation in model species, and therapy of genetic disorders in humans. Sleeping Beauty (SB was the first transposon shown to be capable of gene transfer in vertebrate cells, and recent results confirm that SB supports a full spectrum of genetic engineering including transgenesis, insertional mutagenesis, and therapeutic somatic gene transfer both ex vivo and in vivo. The first clinical application of the SB system will help to validate both the safety and efficacy of this approach. In this review, we describe the major transposon systems currently available (with special emphasis on SB, discuss the various parameters and considerations pertinent to their experimental use, and highlight the state of the art in transposon technology in diverse genetic applications.

The expanding universe of transposon technologies for gene and cell engineering.

Science.gov (United States)

Ivics, Zoltán; Izsvák, Zsuzsanna

2010-12-07

Transposable elements can be viewed as natural DNA transfer vehicles that, similar to integrating viruses, are capable of efficient genomic insertion. The mobility of class II transposable elements (DNA transposons) can be controlled by conditionally providing the transposase component of the transposition reaction. Thus, a DNA of interest (be it a fluorescent marker, a small hairpin (sh)RNA expression cassette, a mutagenic gene trap or a therapeutic gene construct) cloned between the inverted repeat sequences of a transposon-based vector can be used for stable genomic insertion in a regulated and highly efficient manner. This methodological paradigm opened up a number of avenues for genome manipulations in vertebrates, including transgenesis for the generation of transgenic cells in tissue culture, the production of germline transgenic animals for basic and applied research, forward genetic screens for functional gene annotation in model species, and therapy of genetic disorders in humans. Sleeping Beauty (SB) was the first transposon shown to be capable of gene transfer in vertebrate cells, and recent results confirm that SB supports a full spectrum of genetic engineering including transgenesis, insertional mutagenesis, and therapeutic somatic gene transfer both ex vivo and in vivo. The first clinical application of the SB system will help to validate both the safety and efficacy of this approach. In this review, we describe the major transposon systems currently available (with special emphasis on SB), discuss the various parameters and considerations pertinent to their experimental use, and highlight the state of the art in transposon technology in diverse genetic applications.

Complete nucleotide sequence and analysis of two conjugative broad host range plasmids from a marine microbial biofilm.

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Peter Norberg

Full Text Available The complete nucleotide sequence of plasmids pMCBF1 and pMCBF6 was determined and analyzed. pMCBF1 and pMCBF6 form a novel clade within the IncP-1 plasmid family designated IncP-1 ς. The plasmids were exogenously isolated earlier from a marine biofilm. pMCBF1 (62 689 base pairs; bp and pMCBF6 (66 729 bp have identical backbones, but differ in their mercury resistance transposons. pMCBF1 carries Tn5053 and pMCBF6 carries Tn5058. Both are flanked by 5 bp direct repeats, typical of replicative transposition. Both insertions are in the vicinity of a resolvase gene in the backbone, supporting the idea that both transposons are "res-site hunters" that preferably insert close to and use external resolvase functions. The similarity of the backbones indicates recent insertion of the two transposons and the ongoing dynamics of plasmid evolution in marine biofilms. Both plasmids also carry the insertion sequence ISPst1, albeit without flanking repeats. ISPs1is located in an unusual site within the control region of the plasmid. In contrast to most known IncP-1 plasmids the pMCBF1/pMCBF6 backbone has no insert between the replication initiation gene (trfA and the vegetative replication origin (oriV. One pMCBF1/pMCBF6 block of about 2.5 kilo bases (kb has no similarity with known sequences in the databases. Furthermore, insertion of three genes with similarity to the multidrug efflux pump operon mexEF and a gene from the NodT family of the tripartite multi-drug resistance-nodulation-division (RND system in Pseudomonas aeruginosa was found. They do not seem to confer antibiotic resistance to the hosts of pMCBF1/pMCBF6, but the presence of RND on promiscuous plasmids may have serious implications for the spread of antibiotic multi-resistance.

Derivation and characterization of sleeping beauty transposon-mediated porcine induced pluripotent stem cells

DEFF Research Database (Denmark)

Kues, Wilfried A.; Herrmann, Doris; Barg-Kues, Brigitte

2013-01-01

the nonviral Sleeping Beauty transposon system to deliver the reprogramming factors Oct4, Sox2, Klf4, and cMyc. Successful reprogramming to a pluripotent state was indicated by changes in cell morphology and reactivation of the Oct4-EGFP reporter. The transposon-reprogrammed induced pluripotent stem (i......PS) cells showed long-term proliferation in vitro over >40 passages, expressed transcription factors typical of embryonic stem cells, including OCT4, NANOG, SOX2, REX1, ESRRB, DPPA5, and UTF1 and surface markers of pluripotency, including SSEA-1 and TRA-1-60. In vitro differentiation resulted in derivatives......The domestic pig is an important large animal model for preclinical testing of novel cell therapies. Recently, we produced pluripotency reporter pigs in which the Oct4 promoter drives expression of the enhanced green fluorescent protein (EGFP). Here, we reprogrammed Oct4-EGFP fibroblasts employing...

Towards controlled mutagenesis with transposons Ac and Tam3

Energy Technology Data Exchange (ETDEWEB)

Haring, M; Veken, J; Windrich, R; Kneppers, T; Rommens, C; Nijkamp, H J.J.; Hille, J [Department of Genetics, Free University, Amsterdam (Netherlands)

1990-01-01

Full text: The discovery of mobile genetic elements in plants has permitted the use of these transposons for insertional mutagenesis. This applies so far only to Zea mays and Antirrhinum majus, because other plant transposable elements have not been characterised so thoroughly at the genetic and the molecular level. To establish whether transposons (Ac from maize and Tam3 from Antirrhinum) remain mobile in heterologous hosts, either in somatic tissue or after meiosis, a phenotypic assay system for transposition was developed. The separation of the two transposition functions will allow controlled mutagenesis of plant genes. Our results indicate that both transposable elements remain active in heterologous hosts. (author)

The OmpA-like protein Loa22 is essential for leptospiral virulence.

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Paula Ristow

2007-07-01

Full Text Available Pathogenic mechanisms of Leptospira interrogans, the causal agent of leptospirosis, remain largely unknown. This is mainly due to the lack of tools for genetic manipulations of pathogenic species. In this study, we characterized a mutant obtained by insertion of the transposon Himar1 into a gene encoding a putative lipoprotein, Loa22, which has a predicted OmpA domain based on sequence identity. The resulting mutant did not express Loa22 and was attenuated in virulence in the guinea pig and hamster models of leptospirosis, whereas the genetically complemented strain was restored in Loa22 expression and virulence. Our results show that Loa22 was expressed during host infection and exposed on the cell surface. Loa22 is therefore necessary for virulence of L. interrogans in the animal model and represents, to our knowledge, the first genetically defined virulence factor in Leptospira species.

Transposon-mediated BAC transgenesis in zebrafish and mice

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Sumiyama Kenta

2009-10-01

Full Text Available Abstract Background Bacterial artificial chromosomes (BACs are among the most widely used tools for studies of gene regulation and function in model vertebrates, yet methods for predictable delivery of BAC transgenes to the genome are currently limited. This is because BAC transgenes are usually microinjected as naked DNA into fertilized eggs and are known to integrate as multi-copy concatamers in the genome. Although conventional methods for BAC transgenesis have been very fruitful, complementary methods for generating single copy BAC integrations would be desirable for many applications. Results We took advantage of the precise cut-and-paste behavior of a natural transposon, Tol2, to develop a new method for BAC transgenesis. In this new method, the minimal sequences of the Tol2 transposon were used to deliver precisely single copies of a ~70 kb BAC transgene to the zebrafish and mouse genomes. We mapped the BAC insertion sites in the genome by standard PCR methods and confirmed transposase-mediated integrations. Conclusion The Tol2 transposon has a surprisingly large cargo capacity that can be harnessed for BAC transgenesis. The precise delivery of single-copy BAC transgenes by Tol2 represents a useful complement to conventional BAC transgenesis, and could aid greatly in the production of transgenic fish and mice for genomics projects, especially those in which single-copy integrations are desired.

Random transposon mutagenesis of the Saccharopolyspora erythraea genome reveals additional genes influencing erythromycin biosynthesis

Science.gov (United States)

Fedashchin, Andrij; Cernota, William H.; Gonzalez, Melissa C.; Leach, Benjamin I.; Kwan, Noelle; Wesley, Roy K.; Weber, J. Mark

2015-01-01

A single cycle of strain improvement was performed in Saccharopolyspora erythraea mutB and 15 genotypes influencing erythromycin production were found. Genotypes generated by transposon mutagenesis appeared in the screen at a frequency of ∼3%. Mutations affecting central metabolism and regulatory genes were found, as well as hydrolases, peptidases, glycosyl transferases and unknown genes. Only one mutant retained high erythromycin production when scaled-up from micro-agar plug fermentations to shake flasks. This mutant had a knockout of the cwh1 gene (SACE_1598), encoding a cell-wall-associated hydrolase. The cwh1 knockout produced visible growth and morphological defects on solid medium. This study demonstrated that random transposon mutagenesis uncovers strain improvement-related genes potentially useful for strain engineering. PMID:26468041

Restorer-of-Fertility Mutations Recovered in Transposon-Active Lines of S Male-Sterile Maize

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Susan Gabay-Laughnan

2018-01-01

Full Text Available Mitochondria execute key pathways of central metabolism and serve as cellular sensing and signaling entities, functions that depend upon interactions between mitochondrial and nuclear genetic systems. This is exemplified in cytoplasmic male sterility type S (CMS-S of Zea mays, where novel mitochondrial open reading frames are associated with a pollen collapse phenotype, but nuclear restorer-of-fertility (restorer mutations rescue pollen function. To better understand these genetic interactions, we screened Activator-Dissociation (Ac-Ds, Enhancer/Suppressor-mutator (En/Spm, and Mutator (Mu transposon-active CMS-S stocks to recover new restorer mutants. The frequency of restorer mutations increased in transposon-active stocks compared to transposon-inactive stocks, but most mutants recovered from Ac-Ds and En/Spm stocks were unstable, reverting upon backcrossing to CMS-S inbred lines. However, 10 independent restorer mutations recovered from CMS-S Mu transposon stocks were stable upon backcrossing. Many restorer mutations condition seed-lethal phenotypes that provide a convenient test for allelism. Eight such mutants recovered in this study included one pair of allelic mutations that were also allelic to the previously described rfl2-1 mutant. Targeted analysis of mitochondrial proteins by immunoblot identified two features that consistently distinguished restored CMS-S pollen from comparably staged, normal-cytoplasm, nonmutant pollen: increased abundance of nuclear-encoded alternative oxidase relative to mitochondria-encoded cytochrome oxidase and decreased abundance of mitochondria-encoded ATP synthase subunit 1 compared to nuclear-encoded ATP synthase subunit 2. CMS-S restorer mutants thus revealed a metabolic plasticity in maize pollen, and further study of these mutants will provide new insights into mitochondrial functions that are critical to pollen and seed development.

The maternal-effect, selfish genetic element Medea is associated with a composite Tc1 transposon.

Science.gov (United States)

Lorenzen, Marcé D; Gnirke, Andreas; Margolis, Jonathan; Garnes, Jeffrey; Campbell, Margie; Stuart, Jeffrey J; Aggarwal, Rajat; Richards, Stephen; Park, Yoonseong; Beeman, Richard W

2008-07-22

Maternal-Effect Dominant Embryonic Arrest ("Medea") factors are selfish nuclear elements that combine maternal-lethal and zygotic-rescue activities to gain a postzygotic survival advantage. We show that Medea(1) activity in Tribolium castaneum is associated with a composite Tc1 transposon inserted just downstream of the neurotransmitter reuptake symporter bloated tubules (blot), whose Drosophila ortholog has both maternal and zygotic functions. The 21.5-kb insertion contains defective copies of elongation initiation factor-3, ATP synthase subunit C, and an RNaseD-related gene, as well as a potentially intact copy of a prokaryotic DUF1703 gene. Sequence comparisons suggest that the current distribution of Medea(1) reflects global emanation after a single transpositional event in recent evolutionary time. The Medea system in Tribolium represents an unusual type of intragenomic conflict and could provide a useful vehicle for driving desirable genes into populations.

Genome-Wide Analysis of Transposon and Retroviral Insertions Reveals Preferential Integrations in Regions of DNA Flexibility.

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Vrljicak, Pavle; Tao, Shijie; Varshney, Gaurav K; Quach, Helen Ngoc Bao; Joshi, Adita; LaFave, Matthew C; Burgess, Shawn M; Sampath, Karuna

2016-04-07

DNA transposons and retroviruses are important transgenic tools for genome engineering. An important consideration affecting the choice of transgenic vector is their insertion site preferences. Previous large-scale analyses of Ds transposon integration sites in plants were done on the basis of reporter gene expression or germ-line transmission, making it difficult to discern vertebrate integration preferences. Here, we compare over 1300 Ds transposon integration sites in zebrafish with Tol2 transposon and retroviral integration sites. Genome-wide analysis shows that Ds integration sites in the presence or absence of marker selection are remarkably similar and distributed throughout the genome. No strict motif was found, but a preference for structural features in the target DNA associated with DNA flexibility (Twist, Tilt, Rise, Roll, Shift, and Slide) was observed. Remarkably, this feature is also found in transposon and retroviral integrations in maize and mouse cells. Our findings show that structural features influence the integration of heterologous DNA in genomes, and have implications for targeted genome engineering. Copyright © 2016 Vrljicak et al.

Structural basis of hAT transposon end recognition by Hermes, an octameric DNA transposase from Musca domestica.

Science.gov (United States)

Hickman, Alison B; Ewis, Hosam E; Li, Xianghong; Knapp, Joshua A; Laver, Thomas; Doss, Anna-Louise; Tolun, Gökhan; Steven, Alasdair C; Grishaev, Alexander; Bax, Ad; Atkinson, Peter W; Craig, Nancy L; Dyda, Fred

2014-07-17

Hermes is a member of the hAT transposon superfamily that has active representatives, including McClintock's archetypal Ac mobile genetic element, in many eukaryotic species. The crystal structure of the Hermes transposase-DNA complex reveals that Hermes forms an octameric ring organized as a tetramer of dimers. Although isolated dimers are active in vitro for all the chemical steps of transposition, only octamers are active in vivo. The octamer can provide not only multiple specific DNA-binding domains to recognize repeated subterminal sequences within the transposon ends, which are important for activity, but also multiple nonspecific DNA binding surfaces for target capture. The unusual assembly explains the basis of bipartite DNA recognition at hAT transposon ends, provides a rationale for transposon end asymmetry, and suggests how the avidity provided by multiple sites of interaction could allow a transposase to locate its transposon ends amidst a sea of chromosomal DNA. Copyright © 2014 Elsevier Inc. All rights reserved.

Random mutagenesis of the hyperthermophilic archaeon Pyrococcus furiosus using in vitro mariner transposition and natural transformation.

Science.gov (United States)

Guschinskaya, Natalia; Brunel, Romain; Tourte, Maxime; Lipscomb, Gina L; Adams, Michael W W; Oger, Philippe; Charpentier, Xavier

2016-11-08

Transposition mutagenesis is a powerful tool to identify the function of genes, reveal essential genes and generally to unravel the genetic basis of living organisms. However, transposon-mediated mutagenesis has only been successfully applied to a limited number of archaeal species and has never been reported in Thermococcales. Here, we report random insertion mutagenesis in the hyperthermophilic archaeon Pyrococcus furiosus. The strategy takes advantage of the natural transformability of derivatives of the P. furiosus COM1 strain and of in vitro Mariner-based transposition. A transposon bearing a genetic marker is randomly transposed in vitro in genomic DNA that is then used for natural transformation of P. furiosus. A small-scale transposition reaction routinely generates several hundred and up to two thousands transformants. Southern analysis and sequencing showed that the obtained mutants contain a single and random genomic insertion. Polyploidy has been reported in Thermococcales and P. furiosus is suspected of being polyploid. Yet, about half of the mutants obtained on the first selection are homozygous for the transposon insertion. Two rounds of isolation on selective medium were sufficient to obtain gene conversion in initially heterozygous mutants. This transposition mutagenesis strategy will greatly facilitate functional exploration of the Thermococcales genomes.

TCUP: A novel hAT transposon active in maize tissue culture

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Alan eSmith

2012-01-01

Full Text Available Transposable elements are capable of inducing heritable de novo genetic variation. The sequences capable of reactivation, and environmental factors that induce mobilization, remain poorly defined even in well-studied genomes such as maize. We treated maize tissue culture with the demethylating agent 5-aza-2-deoxcytidine and examined long-term tissue culture lines to discover silenced transposable elements that have the potential to induce heritable genetic variation. Through these screens we have identified a novel low copy number hAT transposon, Tissue Culture Up-Regulated (TCUP, which is transcribed at high levels in long-term maize Black Mexican Sweet (BMS tissue culture and up-regulated in response to treatment with 5-aza-2-deoxycytidine. Analysis of the TIGR Maize Gene Index revealed that this element is the most frequently represented EST from the BMS cell culture library and is not represented in other tissue libraries, which is the basis for its name. A full-length sequence was assembled in inbred B73 that contains the putative functional motifs required for autonomous movement of a hAT transposon. Transposon display detected movement of TCUP in two long-term tissue cultured cell lines of the genotype Hi-II AxB and BMS. This research implicates TCUP as a transposon that is capable of reactivation and which may also be particularly sensitive to the stress of the tissue culture environment. Our findings are consistent with the hypothesis that epigenetic alterations potentiate genomic responses to stress during clonal propagation of plants.

Transposon display supports transpositional activity of P elements in ...

Indian Academy of Sciences (India)

. Abstract. Mobilization of two P element subfamilies (canonical and O-type) from Drosophila sturtevanti and D. saltans was evaluated for copy number and transposition activity using the transposon display (TD) technique. Pairwise distances ...

The Functionality of Minimal PiggyBac Transposons in Mammalian Cells

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Boris Troyanovsky

2016-01-01

Full Text Available Minimal piggyBac vectors are a modified single-plasmid version of the classical piggyBac delivery system that can be used for stable transgene integration. These vectors have a truncated terminal domain in the delivery cassette and thus, integrate significantly less flanking transposon DNA into host cell chromatin than classical piggyBac vectors. Herein, we test various characteristics of this modified transposon. The integration efficiency of minimal piggyBac vectors was inversely related to the size of both the transposon and the entire plasmid, but inserts as large as 15 kb were efficiently integrated. Open and super-coiled vectors demonstrated the same integration efficiency while DNA methylation decreased the integration efficiency and silenced the expression of previously integrated sequences in some cell types. Importantly, the incidence of plasmid backbone integration was not increased above that seen in nontransposon control vectors. In BALB/c mice, we demonstrated prolonged expression of two transgenes (intracellular mCherry and secretable Gaussia luciferase when delivered by the minimal piggyBac that resulted in a more sustained antibody production against the immunogenic luciferase than when delivered by a transient (nontransposon vector plasmid. We conclude that minimal piggyBac vectors are an effective alternative to other integrative systems for stable DNA delivery in vitro and in vivo.

Somatic mutagenesis with a Sleeping Beauty transposon system leads to solid tumor formation in zebrafish.

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Maura McGrail

2011-04-01

Full Text Available Large-scale sequencing of human cancer genomes and mouse transposon-induced tumors has identified a vast number of genes mutated in different cancers. One of the outstanding challenges in this field is to determine which genes, when mutated, contribute to cellular transformation and tumor progression. To identify new and conserved genes that drive tumorigenesis we have developed a novel cancer model in a distantly related vertebrate species, the zebrafish, Danio rerio. The Sleeping Beauty (SB T2/Onc transposon system was adapted for somatic mutagenesis in zebrafish. The carp ß-actin promoter was cloned into T2/Onc to create T2/OncZ. Two transgenic zebrafish lines that contain large concatemers of T2/OncZ were isolated by injection of linear DNA into the zebrafish embryo. The T2/OncZ transposons were mobilized throughout the zebrafish genome from the transgene array by injecting SB11 transposase RNA at the 1-cell stage. Alternatively, the T2/OncZ zebrafish were crossed to a transgenic line that constitutively expresses SB11 transposase. T2/OncZ transposon integration sites were cloned by ligation-mediated PCR and sequenced on a Genome Analyzer II. Between 700-6800 unique integration events in individual fish were mapped to the zebrafish genome. The data show that introduction of transposase by transgene expression or RNA injection results in an even distribution of transposon re-integration events across the zebrafish genome. SB11 mRNA injection resulted in neoplasms in 10% of adult fish at ∼10 months of age. T2/OncZ-induced zebrafish tumors contain many mutated genes in common with human and mouse cancer genes. These analyses validate our mutagenesis approach and provide additional support for the involvement of these genes in human cancers. The zebrafish T2/OncZ cancer model will be useful for identifying novel and conserved genetic drivers of human cancers.

Horizontal gene transfer and mobile genetic elements in marine systems.

Science.gov (United States)

Sobecky, Patricia A; Hazen, Tracy H

2009-01-01

The pool of mobile genetic elements (MGE) in microbial communities consists of viruses, plasmids, and associated elements (insertion sequences, transposons, and integrons) that are either self-transmissible or use mobile plasmids and viruses as vehicles for their dissemination. This mobilome facilitates the horizontal transfer of genes that promote the evolution and adaptation of microbial communities. Efforts to characterize MGEs from microbial populations resident in a variety of ecological habitats have revealed a surprisingly novel and seemingly untapped biodiversity. To better understand the impact of horizontal gene transfer (HGT), as well as the agents that promote HGT in marine ecosystems and to determine whether or not environmental parameters can effect the composition and structure of the mobilome in marine microbial communities, information on the distribution, diversity, and ecological traits of the marine mobilome is presented. In this chapter we discuss recent insights gained from different methodological approaches used to characterize the biodiversity and ecology of MGE in marine environments and their contributions to HGT. In addition, we present case studies that highlight specific HGT examples in coastal, open-ocean, and deep-sea marine ecosystems.

Stable gene transfer of CCR5 and CXCR4 siRNAs by sleeping beauty transposon system to confer HIV-1 resistance

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Akkina Ramesh

2008-07-01

Full Text Available Abstract Background Thus far gene therapy strategies for HIV/AIDS have used either conventional retroviral vectors or lentiviral vectors for gene transfer. Although highly efficient, their use poses a certain degree of risk in terms of viral mediated oncogenesis. Sleeping Beauty (SB transposon system offers a non-viral method of gene transfer to avoid this possible risk. With respect to conferring HIV resistance, stable knock down of HIV-1 coreceptors CCR5 and CXCR4 by the use of lentiviral vector delivered siRNAs has proved to be a promising strategy to protect cells from HIV-1 infection. In the current studies our aim is to evaluate the utility of SB system for stable gene transfer of CCR5 and CXCR4 siRNA genes to derive HIV resistant cells as a first step towards using this system for gene therapy. Results Two well characterized siRNAs against the HIV-1 coreceptors CCR5 and CXCR4 were chosen based on their previous efficacy for the SB transposon gene delivery. The siRNA transgenes were incorporated individually into a modified SB transfer plasmid containing a FACS sortable red fluorescence protein (RFP reporter and a drug selectable neomycin resistance gene. Gene transfer was achieved by co-delivery with a construct expressing a hyperactive transposase (HSB5 into the GHOST-R3/X4/R5 cell line, which expresses the major HIV receptor CD4 and and the co-receptors CCR5 and CXCR4. SB constructs expressing CCR5 or CXCR4 siRNAs were also transfected into MAGI-CCR5 or MAGI-CXCR4 cell lines, respectively. Near complete downregulation of CCR5 and CXCR4 surface expression was observed in transfected cells. During viral challenge with X4-tropic (NL4.3 or R5-tropic (BaL HIV-1 strains, the respective transposed cells showed marked viral resistance. Conclusion SB transposon system can be used to deliver siRNA genes for stable gene transfer. The siRNA genes against HIV-1 coreceptors CCR5 and CXCR4 are able to downregulate the respective cell surface proteins

Rim 2/Hipa CACTA transposon display ; A new genetic marker technique in Oryza species

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Lee Ju

2005-03-01

Full Text Available Abstract Background Transposons constitute the major fractions of repetitive sequences in eukaryotes, and have been crucial in the shaping of current genomes. Transposons are generally divided into two classes according to the mechanism underlying their transposition: RNA intermediate class 1 and DNA intermediate class 2. CACTA is a class 2 transposon superfamily, which is found exclusively in plants. As some transposons, including the CACTA superfamily, are highly abundant in plant species, and their nucleotide sequences are highly conserved within a family, they can be utilized as genetic markers, using a slightly modified version of the conventional AFLP protocol. Rim2 /Hipa is a CACTA transposon family having 16 bp consensus TIR sequences to be present in high copy numbers in rice genome. This research was carried out in order to develop a Rim2/Hipa CACTA-AFLP or Rim2/Hipa CACTA-TD (transposon display, hereafter Rim2/Hipa-TD protocol for the study of genetic markers in map construction and the study of genetic diversity in rice. Results Rim2/Hipa-TD generated ample polymorphic profiles among the different rice accessions, and the amplification profiles were highly reproducible between different thermocyclers and Taq polymerases. These amplification profiles allowed for clear distinction between two different ecotypes, Japonica and Indica, of Oryza sativa. In the analysis of RIL populations, the Rim2/Hipa-TD markers were found to be segregated largely in a dominant manner, although in a few cases, non-parental bands were observed in the segregating populations. Upon linkage analysis, the Rim2/Hipa-TD markers were found to be distributed in the regions proximal to the centromeres of the chromosomes. The distribution of the Rim2/Hipa CACTA elements was surveyed in 15 different Oryza species via Rim2/Hipa-TD. While Rim2/Hipa-TD yielded ample amplification profiles between 100 to 700 bp in the AA diploid Oryza species, other species having BB, CC

Induction of rat liver tumor using the Sleeping Beauty transposon and electroporation.

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Park, June-Shine; Kim, Bae-Hwan; Park, Sung Goo; Jung, Sun Young; Lee, Do Hee; Son, Woo-Chan

2013-05-10

The Sleeping Beauty (SB) transposon system has been receiving much attention as a gene transfer method of choice since it allows permanent gene expression after insertion into the host chromosome. However, low transposition frequency in higher eukaryotes limits its use in commonly-used mammalian species. Researchers have therefore attempted to modify gene delivery and expression to overcome this limitation. In mouse liver, tumor induction using SB introduced by the hydrodynamic method has been successfully accomplished. Liver tumor in rat models using SB could also be of great use; however, dose of DNA, injection volume, rate of injection and achieving back pressure limit the use of the hydrodynamics-based gene delivery. In the present study, we combined the electroporation, a relatively simple and easy gene delivery method, with the SB transposon system and as a result successfully induced tumor in rat liver by directly injecting the c-Myc, HRAS and shp53 genes. The tumor phenotype was determined as a sarcomatoid carcinoma. To our knowledge, this is the first demonstration of induction of tumor in the rat liver using the electroporation-enhanced SB transposon system. Copyright © 2013 Elsevier Inc. All rights reserved.

Genome-Wide Mutagenesis in Borrelia burgdorferi.

Science.gov (United States)

Lin, Tao; Gao, Lihui

2018-01-01

Signature-tagged mutagenesis (STM) is a functional genomics approach to identify bacterial virulence determinants and virulence factors by simultaneously screening multiple mutants in a single host animal, and has been utilized extensively for the study of bacterial pathogenesis, host-pathogen interactions, and spirochete and tick biology. The signature-tagged transposon mutagenesis has been developed to investigate virulence determinants and pathogenesis of Borrelia burgdorferi. Mutants in genes important in virulence are identified by negative selection in which the mutants fail to colonize or disseminate in the animal host and tick vector. STM procedure combined with Luminex Flex ® Map™ technology and next-generation sequencing (e.g., Tn-seq) are the powerful high-throughput tools for the determination of Borrelia burgdorferi virulence determinants. The assessment of multiple tissue sites and two DNA resources at two different time points using Luminex Flex ® Map™ technology provides a robust data set. B. burgdorferi transposon mutant screening indicates that a high proportion of genes are the novel virulence determinants that are required for mouse and tick infection. In this protocol, an effective signature-tagged Himar1-based transposon suicide vector was developed and used to generate a sequence-defined library of nearly 4800 mutants in the infectious B. burgdorferi B31 clone. In STM, signature-tagged suicide vectors are constructed by inserting unique DNA sequences (tags) into the transposable elements. The signature-tagged transposon mutants are generated when transposon suicide vectors are transformed into an infectious B. burgdorferi clone, and the transposable element is transposed into the 5'-TA-3' sequence in the B. burgdorferi genome with the signature tag. The transposon library is created and consists of many sub-libraries, each sub-library has several hundreds of mutants with same tags. A group of mice or ticks are infected with a mixed

Characterization and Diversity of Novel PIF/Harbinger DNA Transposons in Brassica Genomes

International Nuclear Information System (INIS)

Nouroz, F.; Noreen, S.; Harrison, H.

2016-01-01

Among DNA transposons, PIF/Harbinger is most recently identified superfamily characterized by 3 bp target site duplications (TSDs), flanked by 14-45 bp terminal inverted repeats (TIRs) and displaying DDD or DDE domain displaying transposase. Their autonomous elements contain two open reading frames, ORF1 and ORF2 encoding superfamily specific transposase and DNA-binding domain. Harbinger DNA transposons are recently identified in few plants. In present study, computational and molecular approaches were used for the identification of 8 Harbinger transposons, of which only 2 were complete with putative trans posase, while rest 6 lack transposase and are considered as defective or non-autonomous elements. They ranged in size from 0.5-4 kb with 3 bp TSDs, 15-42 bp TIRs and internal AT richregions. The PCR amplification of Brassica Harbinger transposase revealed diversity and ancient nature of these elements. The amplification polymorphism of some non-autonomous Harbingers showed species specific distribution. Phylogenetic analyses of transposase clustered them into two clades (monocot and dicot) and five sub-clades. The Brassica, Arabidopsis and Malustransposase clustered into genera specific sub-clades; although a lot of homology in transposase was observed. The multiple sequence alignment of Brassica and related transposase showed homology in five conserved blocks. The DD/Sub 35/E triad and sequences showed similarity to already known Pong-like or Arabidopsis ATIS12 Harbinger transposase in contrast to other transposase having DD/Sub 47/E or DD/Sub 48/E motifs. The present study will be helpful in the characterization of Harbingers, their structural diversity in related genera and Harbinger based molecular markers for varietal/lines identifications. (author)

Use of mariner transposases for one-step delivery and integration of DNA in prokaryotes and eukaryotes by transfection.

Science.gov (United States)

Trubitsyna, Maryia; Michlewski, Gracjan; Finnegan, David J; Elfick, Alistair; Rosser, Susan J; Richardson, Julia M; French, Christopher E

2017-06-02

Delivery of DNA to cells and its subsequent integration into the host genome is a fundamental task in molecular biology, biotechnology and gene therapy. Here we describe an IP-free one-step method that enables stable genome integration into either prokaryotic or eukaryotic cells. A synthetic mariner transposon is generated by flanking a DNA sequence with short inverted repeats. When purified recombinant Mos1 or Mboumar-9 transposase is co-transfected with transposon-containing plasmid DNA, it penetrates prokaryotic or eukaryotic cells and integrates the target DNA into the genome. In vivo integrations by purified transposase can be achieved by electroporation, chemical transfection or Lipofection of the transposase:DNA mixture, in contrast to other published transposon-based protocols which require electroporation or microinjection. As in other transposome systems, no helper plasmids are required since transposases are not expressed inside the host cells, thus leading to generation of stable cell lines. Since it does not require electroporation or microinjection, this tool has the potential to be applied for automated high-throughput creation of libraries of random integrants for purposes including gene knock-out libraries, screening for optimal integration positions or safe genome locations in different organisms, selection of the highest production of valuable compounds for biotechnology, and sequencing. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

Transposon activation mutagenesis as a screening tool for identifying resistance to cancer therapeutics

International Nuclear Information System (INIS)

Chen, Li; Schmidt, Emmett V; Stuart, Lynda; Ohsumi, Toshiro K; Burgess, Shawn; Varshney, Gaurav K; Dastur, Anahita; Borowsky, Mark; Benes, Cyril; Lacy-Hulbert, Adam

2013-01-01

The development of resistance to chemotherapies represents a significant barrier to successful cancer treatment. Resistance mechanisms are complex, can involve diverse and often unexpected cellular processes, and can vary with both the underlying genetic lesion and the origin or type of tumor. For these reasons developing experimental strategies that could be used to understand, identify and predict mechanisms of resistance in different malignant cells would be a major advance. Here we describe a gain-of-function forward genetic approach for identifying mechanisms of resistance. This approach uses a modified piggyBac transposon to generate libraries of mutagenized cells, each containing transposon insertions that randomly activate nearby gene expression. Genes of interest are identified using next-gen high-throughput sequencing and barcode multiplexing is used to reduce experimental cost. Using this approach we successfully identify genes involved in paclitaxel resistance in a variety of cancer cell lines, including the multidrug transporter ABCB1, a previously identified major paclitaxel resistance gene. Analysis of co-occurring transposons integration sites in single cell clone allows for the identification of genes that might act cooperatively to produce drug resistance a level of information not accessible using RNAi or ORF expression screening approaches. We have developed a powerful pipeline to systematically discover drug resistance in mammalian cells in vitro. This cost-effective approach can be readily applied to different cell lines, to identify canonical or context specific resistance mechanisms. Its ability to probe complex genetic context and non-coding genomic elements as well as cooperative resistance events makes it a good complement to RNAi or ORF expression based screens

Essential Genes for In Vitro Growth of the Endophyte Herbaspirillum seropedicae SmR1 as Revealed by Transposon Insertion Site Sequencing.

Science.gov (United States)

Rosconi, Federico; de Vries, Stefan P W; Baig, Abiyad; Fabiano, Elena; Grant, Andrew J

2016-11-15

The interior of plants contains microorganisms (referred to as endophytes) that are distinct from those present at the root surface or in the surrounding soil. Herbaspirillum seropedicae strain SmR1, belonging to the betaproteobacteria, is an endophyte that colonizes crops, including rice, maize, sugarcane, and sorghum. Different approaches have revealed genes and pathways regulated during the interactions of H. seropedicae with its plant hosts. However, functional genomic analysis of transposon (Tn) mutants has been hampered by the lack of genetic tools. Here we successfully employed a combination of in vivo high-density mariner Tn mutagenesis and targeted Tn insertion site sequencing (Tn-seq) in H. seropedicae SmR1. The analysis of multiple gene-saturating Tn libraries revealed that 395 genes are essential for the growth of H. seropedicae SmR1 in tryptone-yeast extract medium. A comparative analysis with the Database of Essential Genes (DEG) showed that 25 genes are uniquely essential in H. seropedicae SmR1. The Tn mutagenesis protocol developed and the gene-saturating Tn libraries generated will facilitate elucidation of the genetic mechanisms of the H. seropedicae endophytic lifestyle. A focal point in the study of endophytes is the development of effective biofertilizers that could help to reduce the input of agrochemicals in croplands. Besides the ability to promote plant growth, a good biofertilizer should be successful in colonizing its host and competing against the native microbiota. By using a systematic Tn-based gene-inactivation strategy and massively parallel sequencing of Tn insertion sites (Tn-seq), it is possible to study the fitness of thousands of Tn mutants in a single experiment. We have applied the combination of these techniques to the plant-growth-promoting endophyte Herbaspirillum seropedicae SmR1. The Tn mutant libraries generated will enable studies into the genetic mechanisms of H. seropedicae-plant interactions. The approach that we

The SUVR4 histone lysine methyltransferase binds ubiquitin and converts H3K9me1 to H3K9me3 on transposon chromatin in Arabidopsis.

Directory of Open Access Journals (Sweden)

Silje V Veiseth

2011-03-01

Full Text Available Chromatin structure and gene expression are regulated by posttranslational modifications (PTMs on the N-terminal tails of histones. Mono-, di-, or trimethylation of lysine residues by histone lysine methyltransferases (HKMTases can have activating or repressive functions depending on the position and context of the modified lysine. In Arabidopsis, trimethylation of lysine 9 on histone H3 (H3K9me3 is mainly associated with euchromatin and transcribed genes, although low levels of this mark are also detected at transposons and repeat sequences. Besides the evolutionarily conserved SET domain which is responsible for enzyme activity, most HKMTases also contain additional domains which enable them to respond to other PTMs or cellular signals. Here we show that the N-terminal WIYLD domain of the Arabidopsis SUVR4 HKMTase binds ubiquitin and that the SUVR4 product specificity shifts from di- to trimethylation in the presence of free ubiquitin, enabling conversion of H3K9me1 to H3K9me3 in vitro. Chromatin immunoprecipitation and immunocytological analysis showed that SUVR4 in vivo specifically converts H3K9me1 to H3K9me3 at transposons and pseudogenes and has a locus-specific repressive effect on the expression of such elements. Bisulfite sequencing indicates that this repression involves both DNA methylation-dependent and -independent mechanisms. Transcribed genes with high endogenous levels of H3K4me3, H3K9me3, and H2Bub1, but low H3K9me1, are generally unaffected by SUVR4 activity. Our results imply that SUVR4 is involved in the epigenetic defense mechanism by trimethylating H3K9 to suppress potentially harmful transposon activity.

Target Capture during Mos1 Transposition*

Science.gov (United States)

Pflieger, Aude; Jaillet, Jerôme; Petit, Agnès; Augé-Gouillou, Corinne; Renault, Sylvaine

2014-01-01

DNA transposition contributes to genomic plasticity. Target capture is a key step in the transposition process, because it contributes to the selection of new insertion sites. Nothing or little is known about how eukaryotic mariner DNA transposons trigger this step. In the case of Mos1, biochemistry and crystallography have deciphered several inverted terminal repeat-transposase complexes that are intermediates during transposition. However, the target capture complex is still unknown. Here, we show that the preintegration complex (i.e., the excised transposon) is the only complex able to capture a target DNA. Mos1 transposase does not support target commitment, which has been proposed to explain Mos1 random genomic integrations within host genomes. We demonstrate that the TA dinucleotide used as the target is crucial both to target recognition and in the chemistry of the strand transfer reaction. Bent DNA molecules are better targets for the capture when the target DNA is nicked two nucleotides apart from the TA. They improve strand transfer when the target DNA contains a mismatch near the TA dinucleotide. PMID:24269942

One to rule them all

Science.gov (United States)

Bouuaert, Corentin Claeys; Tellier, Michael; Chalmers, Ronald

2014-01-01

The development of transposon-based genome manipulation tools can benefit greatly from understanding transposons’ inherent regulatory mechanisms. The Tc1-mariner transposons, which are being widely used in biotechnological applications, are subject to a self-inhibitory mechanism whereby increasing transposase expression beyond a certain point decreases the rate of transposition. In a recent paper, Liu and Chalmers performed saturating mutagenesis on the highly conserved WVPHEL motif in the mariner-family transposase from the Hsmar1 element. Curiously, they found that the majority of all possible single mutations were hyperactive. Biochemical characterizations of the mutants revealed that the hyperactivity is due to a defect in communication between transposase subunits, which normally regulates transposition by reducing the rate of synapsis. This provides important clues for improving transposon-based tools. However, some WVPHEL mutants also showed features that would be undesirable for most biotechnological applications: they showed uncontrolled DNA cleavage activities and defects in the coordination of cleavage between the two transposon ends. The study illustrates how the knowledge of inhibitory mechanisms can help improve transposon tools but also highlights an important challenge, which is to specifically target a regulatory mechanism without affecting other important functions of the transposase. PMID:24812590

Chemical mutagens, transposons, and transgenes to interrogate gene function in Drosophila melanogaster.

Science.gov (United States)

Venken, Koen J T; Bellen, Hugo J

2014-06-15

The study of genetics, genes, and chromosomal inheritance was initiated by Thomas Morgan in 1910, when the first visible mutations were identified in fruit flies. The field expanded upon the work initiated by Herman Muller in 1926 when he used X-rays to develop the first balancer chromosomes. Today, balancers are still invaluable to maintain mutations and transgenes but the arsenal of tools has expanded vastly and numerous new methods have been developed, many relying on the availability of the genome sequence and transposable elements. Forward genetic screens based on chemical mutagenesis or transposable elements have resulted in the unbiased identification of many novel players involved in processes probed by specific phenotypic assays. Reverse genetic approaches have relied on the availability of a carefully selected set of transposon insertions spread throughout the genome to allow the manipulation of the region in the vicinity of each insertion. Lastly, the ability to transform Drosophila with single copy transgenes using transposons or site-specific integration using the ΦC31 integrase has allowed numerous manipulations, including the ability to create and integrate genomic rescue constructs, generate duplications, RNAi knock-out technology, binary expression systems like the GAL4/UAS system as well as other methods. Here, we will discuss the most useful methodologies to interrogate the fruit fly genome in vivo focusing on chemical mutagenesis, transposons and transgenes. Genome engineering approaches based on nucleases and RNAi technology are discussed in following chapters. Copyright © 2014 Elsevier Inc. All rights reserved.

A large inversion in the linear chromosome of Streptomyces griseus caused by replicative transposition of a new Tn3 family transposon.

Science.gov (United States)

Murata, M; Uchida, T; Yang, Y; Lezhava, A; Kinashi, H

2011-04-01

We have comprehensively analyzed the linear chromosomes of Streptomyces griseus mutants constructed and kept in our laboratory. During this study, macrorestriction analysis of AseI and DraI fragments of mutant 402-2 suggested a large chromosomal inversion. The junctions of chromosomal inversion were cloned and sequenced and compared with the corresponding target sequences in the parent strain 2247. Consequently, a transposon-involved mechanism was revealed. Namely, a transposon originally located at the left target site was replicatively transposed to the right target site in an inverted direction, which generated a second copy and at the same time caused a 2.5-Mb chromosomal inversion. The involved transposon named TnSGR was grouped into a new subfamily of the resolvase-encoding Tn3 family transposons based on its gene organization. At the end, terminal diversity of S. griseus chromosomes is discussed by comparing the sequences of strains 2247 and IFO13350.

The transposon Galileo generates natural chromosomal inversions in Drosophila by ectopic recombination.

Science.gov (United States)

Delprat, Alejandra; Negre, Bàrbara; Puig, Marta; Ruiz, Alfredo

2009-11-18

Transposable elements (TEs) are responsible for the generation of chromosomal inversions in several groups of organisms. However, in Drosophila and other Dipterans, where inversions are abundant both as intraspecific polymorphisms and interspecific fixed differences, the evidence for a role of TEs is scarce. Previous work revealed that the transposon Galileo was involved in the generation of two polymorphic inversions of Drosophila buzzatii. To assess the impact of TEs in Drosophila chromosomal evolution and shed light on the mechanism involved, we isolated and sequenced the two breakpoints of another widespread polymorphic inversion from D. buzzatii, 2z(3). In the non inverted chromosome, the 2z(3) distal breakpoint was located between genes CG2046 and CG10326 whereas the proximal breakpoint lies between two novel genes that we have named Dlh and Mdp. In the inverted chromosome, the analysis of the breakpoint sequences revealed relatively large insertions (2,870-bp and 4,786-bp long) including two copies of the transposon Galileo (subfamily Newton), one at each breakpoint, plus several other TEs. The two Galileo copies: (i) are inserted in opposite orientation; (ii) present exchanged target site duplications; and (iii) are both chimeric. Our observations provide the best evidence gathered so far for the role of TEs in the generation of Drosophila inversions. In addition, they show unequivocally that ectopic recombination is the causative mechanism. The fact that the three polymorphic D. buzzatii inversions investigated so far were generated by the same transposon family is remarkable and is conceivably due to Galileo's unusual structure and current (or recent) transpositional activity.

Characterization of Tn3000, a Transposon Responsible for blaNDM-1 Dissemination among Enterobacteriaceae in Brazil, Nepal, Morocco, and India.

Science.gov (United States)

Campos, Juliana Coutinho; da Silva, Maria José Félix; dos Santos, Paulo Roberto Nascimento; Barros, Elaine Menezes; Pereira, Mayne de Oliveira; Seco, Bruna Mara Silva; Magagnin, Cibele Massotti; Leiroz, Leonardo Kalab; de Oliveira, Théo Gremen Mimary; de Faria-Júnior, Célio; Cerdeira, Louise Teixeira; Barth, Afonso Luís; Sampaio, Suely Carlos Ferreira; Zavascki, Alexandre Prehn; Poirel, Laurent; Sampaio, Jorge Luiz Mello

2015-12-01

In Enterobacteriaceae, the blaNDM genes have been found in many different genetic contexts, and a wide diversity of plasmid scaffolds bearing those genes has been found. In August 2013, we identified NDM-1-producing Escherichia coli and Enterobacter hormaechei strains from a single rectal swab sample from a patient hospitalized in Rio de Janeiro, Brazil, who had no history of travel abroad. Complete DNA sequencing using the Illumina platform and annotation of the two plasmids harboring the blaNDM-1 gene, one from each strain, showed that they belonged to incompatibility groups IncFIIK and IncX3 and harbored a novel transposon named Tn3000. Similar genetic structures have been identified among other isolates in Brazil but also on plasmids from other continents. Our findings suggest that the blaNDM-1 gene may be transmitted by Tn3000 in different parts of the world. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Conditional gene expression in the mouse using a Sleeping Beauty gene-trap transposon

Directory of Open Access Journals (Sweden)

Hackett Perry B

2006-06-01

Full Text Available Abstract Background Insertional mutagenesis techniques with transposable elements have been popular among geneticists studying model organisms from E. coli to Drosophila and, more recently, the mouse. One such element is the Sleeping Beauty (SB transposon that has been shown in several studies to be an effective insertional mutagen in the mouse germline. SB transposon vector studies have employed different functional elements and reporter molecules to disrupt and report the expression of endogenous mouse genes. We sought to generate a transposon system that would be capable of reporting the expression pattern of a mouse gene while allowing for conditional expression of a gene of interest in a tissue- or temporal-specific pattern. Results Here we report the systematic development and testing of a transposon-based gene-trap system incorporating the doxycycline-repressible Tet-Off (tTA system that is capable of activating the expression of genes under control of a Tet response element (TRE promoter. We demonstrate that the gene trap system is fully functional in vitro by introducing the "gene-trap tTA" vector into human cells by transposition and identifying clones that activate expression of a TRE-luciferase transgene in a doxycycline-dependent manner. In transgenic mice, we mobilize gene-trap tTA vectors, discover parameters that can affect germline mobilization rates, and identify candidate gene insertions to demonstrate the in vivo functionality of the vector system. We further demonstrate that the gene-trap can act as a reporter of endogenous gene expression and it can be coupled with bioluminescent imaging to identify genes with tissue-specific expression patterns. Conclusion Akin to the GAL4/UAS system used in the fly, we have made progress developing a tool for mutating and revealing the expression of mouse genes by generating the tTA transactivator in the presence of a secondary TRE-regulated reporter molecule. A vector like the gene

A Mos1 transposase in vivo assay to screen new HIV-1 integrase inhibitors.

Science.gov (United States)

Cancian, Mariana; Loreto, Elgion L S

2018-04-01

The integrase and transposase enzymes of retrovirus and transposons, respectively, share the catalytic DDE domain. In vitro assays showed that inhibitors of HIV-1 integrase generally inhibit the mariner Mos1 transposase. Using a Drosophila strain in which the mobilisation of the mariner element can be quantified by mosaic eyes, we showed that flies maintained in medium containing 210 µM to 4 mM of raltegravir, or 1 or 2 mM of dolutegravir, which are HIV-1 integrase inhibitor used in AIDS treatment, have 23-33% less somatic mobilisation in mosaic eyes when treated with raltegravir and 28-32% when treated with dolutegravir. The gene expression of the mariner transposase gene, estimated by qPCR, is similar among treated and control flies. The results suggest that in vivo assays using Drosophila can be used as a primary screening of inhibitory drugs for transposase and retroviral integrase. The advantages of this assay are that it is easy, quick, cheap and is an in vivo test, meaning that the tested substance has to have been taken in by cells and has arrived at the target site, which is not the case when in vitro assays are applied.

Improvement of DNA transfer frequency and transposon mutagenesis of Erwinia carotovora subsp. betavasculorum.

Science.gov (United States)

Rella, M; Axelrood, P E; Weinhold, A R; Schroth, M N

1989-01-01

The production of antibiotics and their role in microbial competition under natural conditions can be readily studied by the use of transposon mutants. Several antibiotic-producing strains of Erwinia carotovora subsp. betavasculorum were unable to accept foreign DNA. A plasmid delivery system was developed, using ethyl methanesulfonate mutagenesis, which entailed isolating E. carotovora subsp. betavasculorum mutants able to accept foreign DNA and transfer it to other strains. This enabled transposon mutagenesis of a wild-type antibiotic-producing strain of E. carotovora subsp. betavasculorum. Twelve antibiotic-negative mutants were isolated, and one of these showed a reduction in antibiotic production in vitro. Many of these mutants also showed a reduction in their ability to macerate potato tissue. The mutants were classified into four genetic groups on the basis of their genetic and phenotypic characteristics, indicating that several genes are involved in antibiotic biosynthesis by E. carotovora subsp. betavasculorum. PMID:2543291

Novel Aminoglycoside Resistance Transposons and Transposon-Derived Circular Forms Detected in Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates

Science.gov (United States)

Dwibedi, Chinmay Kumar; Sjöström, Karin; Edquist, Petra; Wai, Sun Nyunt; Uhlin, Bernt Eric

2016-01-01

Acinetobacter baumannii has emerged as an important opportunistic pathogen equipped with a growing number of antibiotic resistance genes. Our study investigated the molecular epidemiology and antibiotic resistance features of 28 consecutive carbapenem-resistant clinical isolates of A. baumannii collected throughout Sweden in 2012 and 2013. The isolates mainly belonged to clonal complexes (CCs) with an extensive international distribution, such as CC2 (n = 16) and CC25 (n = 7). Resistance to carbapenems was related to blaOXA-23 (20 isolates), blaOXA-24/40-like (6 isolates), blaOXA-467 (1 isolate), and ISAba1-blaOXA-69 (1 isolate). Ceftazidime resistance was associated with blaPER-7 in the CC25 isolates. Two classical point mutations were responsible for resistance to quinolones in all the isolates. Isolates with high levels of resistance to aminoglycosides carried the 16S rRNA methylase armA gene. The isolates also carried a variety of genes encoding aminoglycoside-modifying enzymes. Several novel structures involved in aminoglycoside resistance were identified, including Tn6279, ΔTn6279, Ab-ST3-aadB, and different assemblies of Tn6020 and TnaphA6. Importantly, a number of circular forms related to the IS26 or ISAba125 composite transposons were detected. The frequent occurrence of these circular forms in the populations of several isolates indicates a potential role of these circular forms in the dissemination of antibiotic resistance genes. PMID:26824943

Using PATIMDB to create bacterial transposon insertion mutant libraries.

Science.gov (United States)

Urbach, Jonathan M; Wei, Tao; Liberati, Nicole; Grenfell-Lee, Daniel; Villanueva, Jacinto; Wu, Gang; Ausubel, Frederick M

2009-04-01

PATIMDB is a software package for facilitating the generation of transposon mutant insertion libraries. The software has two main functions: process tracking and automated sequence analysis. The process tracking function specifically includes recording the status and fates of multiwell plates and samples in various stages of library construction. Automated sequence analysis refers specifically to the pipeline of sequence analysis starting with ABI files from a sequencing facility and ending with insertion location identifications. The protocols in this unit describe installation and use of PATIMDB software.

The Tol2 transposon system mediates the genetic engineering of T-cells with CD19-specific chimeric antigen receptors for B-cell malignancies.

Science.gov (United States)

Tsukahara, T; Iwase, N; Kawakami, K; Iwasaki, M; Yamamoto, C; Ohmine, K; Uchibori, R; Teruya, T; Ido, H; Saga, Y; Urabe, M; Mizukami, H; Kume, A; Nakamura, M; Brentjens, R; Ozawa, K

2015-02-01

Engineered T-cell therapy using a CD19-specific chimeric antigen receptor (CD19-CAR) is a promising strategy for the treatment of advanced B-cell malignancies. Gene transfer of CARs to T-cells has widely relied on retroviral vectors, but transposon-based gene transfer has recently emerged as a suitable nonviral method to mediate stable transgene expression. The advantages of transposon vectors compared with viral vectors include their simplicity and cost-effectiveness. We used the Tol2 transposon system to stably transfer CD19-CAR into human T-cells. Normal human peripheral blood lymphocytes were co-nucleofected with the Tol2 transposon donor plasmid carrying CD19-CAR and the transposase expression plasmid and were selectively propagated on NIH3T3 cells expressing human CD19. Expanded CD3(+) T-cells with stable and high-level transgene expression (~95%) produced interferon-γ upon stimulation with CD19 and specifically lysed Raji cells, a CD19(+) human B-cell lymphoma cell line. Adoptive transfer of these T-cells suppressed tumor progression in Raji tumor-bearing Rag2(-/-)γc(-/-) immunodeficient mice compared with control mice. These results demonstrate that the Tol2 transposon system could be used to express CD19-CAR in genetically engineered T-cells for the treatment of refractory B-cell malignancies.

MiMIC: a highly versatile transposon insertion resource for engineering Drosophila melanogaster genes

Science.gov (United States)

Venken, Koen J. T.; Schulze, Karen L.; Haelterman, Nele A.; Pan, Hongling; He, Yuchun; Evans-Holm, Martha; Carlson, Joseph W.; Levis, Robert W.; Spradling, Allan C.; Hoskins, Roger A.; Bellen, Hugo J.

2011-01-01

We demonstrate the versatility of a collection of insertions of the transposon Minos mediated integration cassette (MiMIC), in Drosophila melanogaster. MiMIC contains a gene-trap cassette and the yellow+ marker flanked by two inverted bacteriophage ΦC31 attP sites. MiMIC integrates almost at random in the genome to create sites for DNA manipulation. The attP sites allow the replacement of the intervening sequence of the transposon with any other sequence through recombinase mediated cassette exchange (RMCE). We can revert insertions that function as gene traps and cause mutant phenotypes to wild type by RMCE and modify insertions to control GAL4 or QF overexpression systems or perform lineage analysis using the Flp system. Insertions within coding introns can be exchanged with protein-tag cassettes to create fusion proteins to follow protein expression and perform biochemical experiments. The applications of MiMIC vastly extend the Drosophila melanogaster toolkit. PMID:21985007

1st Advanced Marine Renewable Energy Instrumentation Experts Workshop

Energy Technology Data Exchange (ETDEWEB)

none,

2011-10-01

The U.S. marine energy industry is actively pursuing development of offshore wind and marine hydrokinetic (MHK) energy systems. Experience in the wind energy sector demonstrates that new technology development requires thorough measurement and characterization of the environmental conditions prevalent at installation sites and of technology operating in the field. Presently, there are no turn-key instrumentation system solutions that meet the measurement needs of the marine energy industry. The 1st Advanced Marine Renewable Energy Instrumentation Experts Workshop brought together technical experts from government laboratories, academia, and industry representatives from marine energy, wind, offshore oil and gas, and instrumentation developers to present and discuss the instrumentation needs of the marine energy industry. The goals of the meeting were to: 1. Share the latest relevant knowledge among technical experts; 2. Review relevant state-of-the-art field measurement technologies and methods; 3. Review lessons learned from recent field deployments; 4. Identify synergies across different industries; 5. Identify gaps between existing and needed instrumentation capabilities; 6. Understand who are the leading experts; 7. Provide a forum where stakeholders from the marine energy industry could provide substantive input in the development of new marine energy field deployable instrumentation packages.

Transposable elements in fish chromosomes: a study in the marine cobia species.

Science.gov (United States)

Costa, G W W F; Cioffi, M B; Bertollo, L A C; Molina, W F

2013-01-01

Rachycentron canadum, a unique representative of the Rachycentridae family, has been the subject of considerable biotechnological interest due to its potential use in marine fish farming. This species has undergone extensive research concerning the location of genes and multigene families on its chromosomes. Although most of the genome of some organisms is composed of repeated DNA sequences, aspects of the origin and dispersion of these elements are still largely unknown. The physical mapping of repetitive sequences on the chromosomes of R. canadum proved to be relevant for evolutionary and applied purposes. Therefore, here, we present the mapping by fluorescence in situ hybridization of the transposable element (TE) Tol2, the non-LTR retrotransposons Rex1 and Rex3, together with the 18S and 5S rRNA genes in the chromosome of this species. The Tol2 TE, belonging to the family of hAT transposons, is homogeneously distributed in the euchromatic regions of the chromosomes but with huge colocalization with the 18S rDNA sites. The hybridization signals for Rex1 and Rex3 revealed a semi-arbitrary distribution pattern, presenting differentiated dispersion in euchromatic and heterochromatic regions. Rex1 elements are associated preferentially in heterochromatic regions, while Rex3 shows a scarce distribution in the euchromatic regions of the chromosomes. The colocalization of TEs with 18S and 5S rDNA revealed complex chromosomal regions of repetitive sequences. In addition, the nonpreferential distribution of Rex1 and Rex3 in all heterochromatic regions, as well as the preferential distribution of the Tol2 transposon associated with 18S rDNA sequences, reveals a distinct pattern of organization of TEs in the genome of this species. A heterogeneous chromosomal colonization of TEs may confer different evolutionary rates to the heterochromatic regions of this species.

Transposon mutations in the flagella biosynthetic pathway of the solvent-tolerant Pseudomonas putida S12 result in a decreased expression of solvent efflux genes

NARCIS (Netherlands)

Kieboom, J; Bruinenberg, R; Keizer-Gunnink, [No Value; de Bont, JAM

2001-01-01

Fourteen solvent-sensitive transposon mutants were generated from the solvent-tolerant Pseudomonas putida strain S12 by applying the TnMOD-KmO mutagenesis system. These mutants were unable to grow in the presence of octanol and toluene. By cloning the region flanking the transposon insertion point a

Random mutagenesis in Corynebacterium glutamicum ATCC 13032 using an IS6100-based transposon vector identified the last unknown gene in the histidine biosynthesis pathway

Directory of Open Access Journals (Sweden)

Gaigalat Lars

2006-08-01

Full Text Available Abstract Background Corynebacterium glutamicum, a Gram-positive bacterium of the class Actinobacteria, is an industrially relevant producer of amino acids. Several methods for the targeted genetic manipulation of this organism and rational strain improvement have been developed. An efficient transposon mutagenesis system for the completely sequenced type strain ATCC 13032 would significantly advance functional genome analysis in this bacterium. Results A comprehensive transposon mutant library comprising 10,080 independent clones was constructed by electrotransformation of the restriction-deficient derivative of strain ATCC 13032, C. glutamicum RES167, with an IS6100-containing non-replicative plasmid. Transposon mutants had stable cointegrates between the transposon vector and the chromosome. Altogether 172 transposon integration sites have been determined by sequencing of the chromosomal inserts, revealing that each integration occurred at a different locus. Statistical target site analyses revealed an apparent absence of a target site preference. From the library, auxotrophic mutants were obtained with a frequency of 2.9%. By auxanography analyses nearly two thirds of the auxotrophs were further characterized, including mutants with single, double and alternative nutritional requirements. In most cases the nutritional requirement observed could be correlated to the annotation of the mutated gene involved in the biosynthesis of an amino acid, a nucleotide or a vitamin. One notable exception was a clone mutagenized by transposition into the gene cg0910, which exhibited an auxotrophy for histidine. The protein sequence deduced from cg0910 showed high sequence similarities to inositol-1(or 4-monophosphatases (EC 3.1.3.25. Subsequent genetic deletion of cg0910 delivered the same histidine-auxotrophic phenotype. Genetic complementation of the mutants as well as supplementation by histidinol suggests that cg0910 encodes the hitherto unknown

33 CFR 1.05-5 - Marine Safety and Security Council.

Science.gov (United States)

2010-07-01

... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Marine Safety and Security... SECURITY GENERAL GENERAL PROVISIONS Rulemaking § 1.05-5 Marine Safety and Security Council. The Marine Safety and Security Council, composed of senior Coast Guard officials, acts as policy advisor to the...

Marine environment news. Vol. 1, no. 1

International Nuclear Information System (INIS)

2003-09-01

This is the first issue of the IAEA's Marine Environment Newsletter which is hoped to inform Member States, research partners, visitors and other stakeholders of highlights of the marine projects, surveys, hot issues, discoveries and training programmes being delivered by the IAEA's Marine Environment Laboratory (MEL) in Monaco. In this issue the mission of the MEL and its various activities are presented

Transcription and somatic transposition of the maize En/Spm transposon system in rice

NARCIS (Netherlands)

Greco, R.; Ouwerkerk, P.B.F.; Taal, A.J.C.; Sallaud, C.; Guiderdoni, E.; Meijer, A.H.; Hoge, J.H.C.; Pereira, A.B.

2004-01-01

Transposition of the maize En/Spm system in rice was investigated using a two-component construct consisting of an immobilised transposase source driven by the CaMV 35S-promoter, and a modified I/dSpm transposon. Mobilization of I/dSpm in somatic sectors was demonstrated by sequencing of excision

TAPDANCE: An automated tool to identify and annotate transposon insertion CISs and associations between CISs from next generation sequence data

Directory of Open Access Journals (Sweden)

Sarver Aaron L

2012-06-01

Full Text Available Abstract Background Next generation sequencing approaches applied to the analyses of transposon insertion junction fragments generated in high throughput forward genetic screens has created the need for clear informatics and statistical approaches to deal with the massive amount of data currently being generated. Previous approaches utilized to 1 map junction fragments within the genome and 2 identify Common Insertion Sites (CISs within the genome are not practical due to the volume of data generated by current sequencing technologies. Previous approaches applied to this problem also required significant manual annotation. Results We describe Transposon Annotation Poisson Distribution Association Network Connectivity Environment (TAPDANCE software, which automates the identification of CISs within transposon junction fragment insertion data. Starting with barcoded sequence data, the software identifies and trims sequences and maps putative genomic sequence to a reference genome using the bowtie short read mapper. Poisson distribution statistics are then applied to assess and rank genomic regions showing significant enrichment for transposon insertion. Novel methods of counting insertions are used to ensure that the results presented have the expected characteristics of informative CISs. A persistent mySQL database is generated and utilized to keep track of sequences, mappings and common insertion sites. Additionally, associations between phenotypes and CISs are also identified using Fisher’s exact test with multiple testing correction. In a case study using previously published data we show that the TAPDANCE software identifies CISs as previously described, prioritizes them based on p-value, allows holistic visualization of the data within genome browser software and identifies relationships present in the structure of the data. Conclusions The TAPDANCE process is fully automated, performs similarly to previous labor intensive approaches

TED, an autonomous and rare maize transposon of the mutator superfamily with a high gametophytic excision frequency.

Science.gov (United States)

Li, Yubin; Harris, Linda; Dooner, Hugo K

2013-09-01

Mutator (Mu) elements, one of the most diverse superfamilies of DNA transposons, are found in all eukaryotic kingdoms, but are particularly numerous in plants. Most of the present knowledge on the transposition behavior of this superfamily comes from studies of the maize (Zea mays) Mu elements, whose transposition is mediated by the autonomous Mutator-Don Robertson (MuDR) element. Here, we describe the maize element TED (for Transposon Ellen Dempsey), an autonomous cousin that differs significantly from MuDR. Element excision and reinsertion appear to require both proteins encoded by MuDR, but only the single protein encoded by TED. Germinal excisions, rare with MuDR, are common with TED, but arise in one of the mitotic divisions of the gametophyte, rather than at meiosis. Instead, transposition-deficient elements arise at meiosis, suggesting that the double-strand breaks produced by element excision are repaired differently in mitosis and meiosis. Unlike MuDR, TED is a very low-copy transposon whose number and activity do not undergo dramatic changes upon inbreeding or outcrossing. Like MuDR, TED transposes mostly to unlinked sites and can form circular transposition products. Sequences closer to TED than to MuDR were detected only in the grasses, suggesting a rather recent evolutionary split from a common ancestor.

piggyBac transposon somatic mutagenesis with an activated reporter and tracker (PB-SMART for genetic screens in mice.

Directory of Open Access Journals (Sweden)

Sean F Landrette

Full Text Available Somatic forward genetic screens have the power to interrogate thousands of genes in a single animal. Retroviral and transposon mutagenesis systems in mice have been designed and deployed in somatic tissues for surveying hematopoietic and solid tumor formation. In the context of cancer, the ability to visually mark mutant cells would present tremendous advantages for identifying tumor formation, monitoring tumor growth over time, and tracking tumor infiltrations and metastases into wild-type tissues. Furthermore, locating mutant clones is a prerequisite for screening and analyzing most other somatic phenotypes. For this purpose, we developed a system using the piggyBac (PB transposon for somatic mutagenesis with an activated reporter and tracker, called PB-SMART. The PB-SMART mouse genetic screening system can simultaneously induce somatic mutations and mark mutated cells using bioluminescence or fluorescence. The marking of mutant cells enable analyses that are not possible with current somatic mutagenesis systems, such as tracking cell proliferation and tumor growth, detecting tumor cell infiltrations, and reporting tissue mutagenesis levels by a simple ex vivo visual readout. We demonstrate that PB-SMART is highly mutagenic, capable of tumor induction with low copy transposons, which facilitates the mapping and identification of causative insertions. We further integrated a conditional transposase with the PB-SMART system, permitting tissue-specific mutagenesis with a single cross to any available Cre line. Targeting the germline, the system could also be used to conduct F1 screens. With these features, PB-SMART provides an integrated platform for individual investigators to harness the power of somatic mutagenesis and phenotypic screens to decipher the genetic basis of mammalian biology and disease.

Glutathione provides a source of cysteine essential for intracellular multiplication of Francisella tularensis.

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Khaled Alkhuder

2009-01-01

Full Text Available Francisella tularensis is a highly infectious bacterium causing the zoonotic disease tularemia. Its ability to multiply and survive in macrophages is critical for its virulence. By screening a bank of HimarFT transposon mutants of the F. tularensis live vaccine strain (LVS to isolate intracellular growth-deficient mutants, we selected one mutant in a gene encoding a putative gamma-glutamyl transpeptidase (GGT. This gene (FTL_0766 was hence designated ggt. The mutant strain showed impaired intracellular multiplication and was strongly attenuated for virulence in mice. Here we present evidence that the GGT activity of F. tularensis allows utilization of glutathione (GSH, gamma-glutamyl-cysteinyl-glycine and gamma-glutamyl-cysteine dipeptide as cysteine sources to ensure intracellular growth. This is the first demonstration of the essential role of a nutrient acquisition system in the intracellular multiplication of F. tularensis. GSH is the most abundant source of cysteine in the host cytosol. Thus, the capacity this intracellular bacterial pathogen has evolved to utilize the available GSH, as a source of cysteine in the host cytosol, constitutes a paradigm of bacteria-host adaptation.

A transposon mutant library of Bacillus cereus ATCC 10987 reveals novel genes required for biofilm formation and implicates motility as an important factor for pellicle-biofilm formation.

Science.gov (United States)

Okshevsky, Mira; Louw, Matilde Greve; Lamela, Elena Otero; Nilsson, Martin; Tolker-Nielsen, Tim; Meyer, Rikke Louise

2018-04-01

Bacillus cereus is one of the most common opportunistic pathogens causing foodborne illness, as well as a common source of contamination in the dairy industry. B. cereus can form robust biofilms on food processing surfaces, resulting in food contamination due to shedding of cells and spores. Despite the medical and industrial relevance of this species, the genetic basis of biofilm formation in B. cereus is not well studied. In order to identify genes required for biofilm formation in this bacterium, we created a library of 5000 +  transposon mutants of the biofilm-forming strain B. cereusATCC 10987, using an unbiased mariner transposon approach. The mutant library was screened for the ability to form a pellicle biofilm at the air-media interface, as well as a submerged biofilm at the solid-media interface. A total of 91 genes were identified as essential for biofilm formation. These genes encode functions such as chemotaxis, amino acid metabolism and cellular repair mechanisms, and include numerous genes not previously known to be required for biofilm formation. Although the majority of disrupted genes are not directly responsible for motility, further investigations revealed that the vast majority of the biofilm-deficient mutants were also motility impaired. This observation implicates motility as a pivotal factor in the formation of a biofilm by B. cereus. These results expand our knowledge of the fundamental molecular mechanisms of biofilm formation by B. cereus. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

A TALE of transposition: Tn3-like transposons play a major role in the spread of pathogenicity determinants of Xanthomonas citri and other xanthomonads.

Science.gov (United States)

Ferreira, Rafael Marini; de Oliveira, Amanda Carolina P; Moreira, Leandro M; Belasque, José; Gourbeyre, Edith; Siguier, Patricia; Ferro, Maria Inês T; Ferro, Jesus A; Chandler, Michael; Varani, Alessandro M

2015-02-17

Members of the genus Xanthomonas are among the most important phytopathogens. A key feature of Xanthomonas pathogenesis is the translocation of type III secretion system (T3SS) effector proteins (T3SEs) into the plant target cells via a T3SS. Several T3SEs and a murein lytic transglycosylase gene (mlt, required for citrus canker symptoms) are found associated with three transposition-related genes in Xanthomonas citri plasmid pXAC64. These are flanked by short inverted repeats (IRs). The region was identified as a transposon, TnXax1, with typical Tn3 family features, including a transposase and two recombination genes. Two 14-bp palindromic sequences within a 193-bp potential resolution site occur between the recombination genes. Additional derivatives carrying different T3SEs and other passenger genes occur in different Xanthomonas species. The T3SEs include transcription activator-like effectors (TALEs). Certain TALEs are flanked by the same IRs as found in TnXax1 to form mobile insertion cassettes (MICs), suggesting that they may be transmitted horizontally. A significant number of MICs carrying other passenger genes (including a number of TALE genes) were also identified, flanked by the same TnXax1 IRs and delimited by 5-bp target site duplications. We conclude that a large fraction of T3SEs, including individual TALEs and potential pathogenicity determinants, have spread by transposition and that TnXax1, which exhibits all of the essential characteristics of a functional transposon, may be involved in driving MIC transposition. We also propose that TALE genes may diversify by fork slippage during the replicative Tn3 family transposition. These mechanisms may play a crucial role in the emergence of Xanthomonas pathogenicity. Xanthomonas genomes carry many insertion sequences (IS) and transposons, which play an important role in their evolution and architecture. This study reveals a key relationship between transposons and pathogenicity determinants in

TED, an Autonomous and Rare Maize Transposon of the Mutator Superfamily with a High Gametophytic Excision Frequency[W

Science.gov (United States)

Li, Yubin; Harris, Linda; Dooner, Hugo K.

2013-01-01

Mutator (Mu) elements, one of the most diverse superfamilies of DNA transposons, are found in all eukaryotic kingdoms, but are particularly numerous in plants. Most of the present knowledge on the transposition behavior of this superfamily comes from studies of the maize (Zea mays) Mu elements, whose transposition is mediated by the autonomous Mutator-Don Robertson (MuDR) element. Here, we describe the maize element TED (for Transposon Ellen Dempsey), an autonomous cousin that differs significantly from MuDR. Element excision and reinsertion appear to require both proteins encoded by MuDR, but only the single protein encoded by TED. Germinal excisions, rare with MuDR, are common with TED, but arise in one of the mitotic divisions of the gametophyte, rather than at meiosis. Instead, transposition-deficient elements arise at meiosis, suggesting that the double-strand breaks produced by element excision are repaired differently in mitosis and meiosis. Unlike MuDR, TED is a very low-copy transposon whose number and activity do not undergo dramatic changes upon inbreeding or outcrossing. Like MuDR, TED transposes mostly to unlinked sites and can form circular transposition products. Sequences closer to TED than to MuDR were detected only in the grasses, suggesting a rather recent evolutionary split from a common ancestor. PMID:24038653

Transposon mutagenesis of probiotic Lactobacillus casei identifies asnH, an asparagine synthetase gene involved in its immune-activating capacity.

Science.gov (United States)

Ito, Masahiro; Kim, Yun-Gi; Tsuji, Hirokazu; Takahashi, Takuya; Kiwaki, Mayumi; Nomoto, Koji; Danbara, Hirofumi; Okada, Nobuhiko

2014-01-01

Lactobacillus casei ATCC 27139 enhances host innate immunity, and the J1 phage-resistant mutants of this strain lose the activity. A transposon insertion mutant library of L. casei ATCC 27139 was constructed, and nine J1 phage-resistant mutants out of them were obtained. Cloning and sequencing analyses identified three independent genes that were disrupted by insertion of the transposon element: asnH, encoding asparagine synthetase, and dnaJ and dnaK, encoding the molecular chaperones DnaJ and DnaK, respectively. Using an in vivo mouse model of Listeria infection, only asnH mutant showed deficiency in their ability to enhance host innate immunity, and complementation of the mutation by introduction of the wild-type asnH in the mutant strain recovered the immuno-augmenting activity. AsnH protein exhibited asparagine synthetase activity when the lysozyme-treated cell wall extracts of L. casei ATCC 27139 was added as substrate. The asnH mutants lost the thick and rigid peptidoglycan features that are characteristic to the wild-type cells, indicating that AsnH of L. casei is involved in peptidoglycan biosynthesis. These results indicate that asnH is required for the construction of the peptidoglycan composition involved in the immune-activating capacity of L. casei ATCC 27139.

Transposon mutagenesis of probiotic Lactobacillus casei identifies asnH, an asparagine synthetase gene involved in its immune-activating capacity.

Directory of Open Access Journals (Sweden)

Masahiro Ito

Full Text Available Lactobacillus casei ATCC 27139 enhances host innate immunity, and the J1 phage-resistant mutants of this strain lose the activity. A transposon insertion mutant library of L. casei ATCC 27139 was constructed, and nine J1 phage-resistant mutants out of them were obtained. Cloning and sequencing analyses identified three independent genes that were disrupted by insertion of the transposon element: asnH, encoding asparagine synthetase, and dnaJ and dnaK, encoding the molecular chaperones DnaJ and DnaK, respectively. Using an in vivo mouse model of Listeria infection, only asnH mutant showed deficiency in their ability to enhance host innate immunity, and complementation of the mutation by introduction of the wild-type asnH in the mutant strain recovered the immuno-augmenting activity. AsnH protein exhibited asparagine synthetase activity when the lysozyme-treated cell wall extracts of L. casei ATCC 27139 was added as substrate. The asnH mutants lost the thick and rigid peptidoglycan features that are characteristic to the wild-type cells, indicating that AsnH of L. casei is involved in peptidoglycan biosynthesis. These results indicate that asnH is required for the construction of the peptidoglycan composition involved in the immune-activating capacity of L. casei ATCC 27139.

Tumor-directed gene therapy in mice using a composite nonviral gene delivery system consisting of the piggyBac transposon and polyethylenimine

International Nuclear Information System (INIS)

Kang, Yu; Zhang, Xiaoyan; Jiang, Wei; Wu, Chaoqun; Chen, Chunmei; Zheng, Yufang; Gu, Jianren; Xu, Congjian

2009-01-01

Compared with viral vectors, nonviral vectors are less immunogenic, more stable, safer and easier to replication for application in cancer gene therapy. However, nonviral gene delivery system has not been extensively used because of the low transfection efficiency and the short transgene expression, especially in vivo. It is desirable to develop a nonviral gene delivery system that can support stable genomic integration and persistent gene expression in vivo. Here, we used a composite nonviral gene delivery system consisting of the piggyBac (PB) transposon and polyethylenimine (PEI) for long-term transgene expression in mouse ovarian tumors. A recombinant plasmid PB [Act-RFP, HSV-tk] encoding both the herpes simplex thymidine kinase (HSV-tk) and the monomeric red fluorescent protein (mRFP1) under PB transposon elements was constructed. This plasmid and the PBase plasmid were injected into ovarian cancer tumor xenografts in mice by in vivo PEI system. The antitumor effects of HSV-tk/ganciclovir (GCV) system were observed after intraperitoneal injection of GCV. Histological analysis and TUNEL assay were performed on the cryostat sections of the tumor tissue. Plasmid construction was confirmed by PCR analysis combined with restrictive enzyme digestion. mRFP1 expression could be visualized three weeks after the last transfection of pPB/TK under fluorescence microscopy. After GCV admission, the tumor volume of PB/TK group was significantly reduced and the tumor inhibitory rate was 81.96% contrasted against the 43.07% in the TK group. Histological analysis showed that there were extensive necrosis and lymphocytes infiltration in the tumor tissue of the PB/TK group but limited in the tissue of control group. TUNEL assays suggested that the transfected cells were undergoing apoptosis after GCV admission in vivo. Our results show that the nonviral gene delivery system coupling PB transposon with PEI can be used as an efficient tool for gene therapy in ovarian cancer

Altered expression of testis-specific genes, piRNAs, and transposons in the silkworm ovary masculinized by a W chromosome mutation

Science.gov (United States)

2012-01-01

Background In the silkworm, Bombyx mori, femaleness is strongly controlled by the female-specific W chromosome. Originally, it was presumed that the W chromosome encodes female-determining gene(s), accordingly called Fem. However, to date, neither Fem nor any protein-coding gene has been identified from the W chromosome. Instead, the W chromosome is occupied with numerous transposon-related sequences. Interestingly, the silkworm W chromosome is a source of female-enriched PIWI-interacting RNAs (piRNAs). piRNAs are small RNAs of 23-30 nucleotides in length, which are required for controlling transposon activity in animal gonads. A recent study has identified a novel mutant silkworm line called KG, whose mutation in the W chromosome causes severe female masculinization. However, the molecular nature of KG line has not been well characterized yet. Results Here we molecularly characterize the KG line. Genomic PCR analyses using currently available W chromosome-specific PCR markers indicated that no large deletion existed in the KG W chromosome. Genetic analyses demonstrated that sib-crosses within the KG line suppressed masculinization. Masculinization reactivated when crossing KG females with wild type males. Importantly, the KG ovaries exhibited a significantly abnormal transcriptome. First, the KG ovaries misexpressed testis-specific genes. Second, a set of female-enriched piRNAs was downregulated in the KG ovaries. Third, several transposons were overexpressed in the KG ovaries. Conclusions Collectively, the mutation in the KG W chromosome causes broadly altered expression of testis-specific genes, piRNAs, and transposons. To our knowledge, this is the first study that describes a W chromosome mutant with such an intriguing phenotype. PMID:22452797

PLE-wu, a new member of piggyBac transposon family from insect, is active in mammalian cells.

Science.gov (United States)

Wu, Chunxiao; Wang, Shu

2014-10-01

piggyBac, a highly active transposon in insect and mammalian cells, is a very useful tool in genome manipulation. A new piggyBac-like element (PLE), named PLE-wu, was identified from a mutant baculovirus cultured in sf9 insect cells. This new transposon is 2931 bp in length and encodes two active forms of transposase, a 708-amino acid-long transposase and a short 576-residue-long transposase translated from a downstream in-frame initiation codon. PLE-wu has asymmetric terminal structures, containing 6-bp inverted terminal repeats, 32-bp imperfect inverted and direct sub-terminal repeats. Similar to piggyBac, PLE-wu exhibits traceless excision activity in both insect and mammalian cells, restoring the original TTAA target sequence upon excision. It also retains the insertion activity in mammalian cells with a plasmid to chromosome transposition rate about 10-fold higher than random integration. Plasmid rescue assays revealed that the TTAA target sequence was duplicated at the junctions of the insertion site. Deletion of the terminal sequences including the sub-terminal repeats decreased the transposition activity of the 708-residue-long transposase, while the transposition activity of the short form of transposase was not affected. With its low sequence similarity to piggyBac, PLE-wu will contribute to the understanding the mechanism of PLE transposition, as well as design of new transposon systems with higher activity. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Improvement of DNA transfer frequency and transposon mutagenesis of Erwinia carotovora subsp. betavasculorum.

OpenAIRE

Rella, M; Axelrood, P E; Weinhold, A R; Schroth, M N

1989-01-01

The production of antibiotics and their role in microbial competition under natural conditions can be readily studied by the use of transposon mutants. Several antibiotic-producing strains of Erwinia carotovora subsp. betavasculorum were unable to accept foreign DNA. A plasmid delivery system was developed, using ethyl methanesulfonate mutagenesis, which entailed isolating E. carotovora subsp. betavasculorum mutants able to accept foreign DNA and transfer it to other strains. This enabled tra...

Global Combat Support System-Marine Corps Logistics Chain Management Increment 1 (GCSS-MC LCM Inc 1)

Science.gov (United States)

2016-03-01

Global Combat Support System – Marine Corps (GCSS-MC) is a portfolio of systems that supports logistics elements of command and control, joint...future Marine Corps logistics systems modernization . GCSS-MC/LCM Increment 1 Capability Release 1 (hereinafter Release 1.1) is the baseline

Fast and efficient Drosophila melanogaster gene knock-ins using MiMIC transposons.

Science.gov (United States)

Vilain, Sven; Vanhauwaert, Roeland; Maes, Ine; Schoovaerts, Nils; Zhou, Lujia; Soukup, Sandra; da Cunha, Raquel; Lauwers, Elsa; Fiers, Mark; Verstreken, Patrik

2014-10-08

Modern molecular genetics studies necessitate the manipulation of genes in their endogenous locus, but most of the current methodologies require an inefficient donor-dependent homologous recombination step to locally modify the genome. Here we describe a methodology to efficiently generate Drosophila knock-in alleles by capitalizing on the availability of numerous genomic MiMIC transposon insertions carrying recombinogenic attP sites. Our methodology entails the efficient PhiC31-mediated integration of a recombination cassette flanked by unique I-SceI and/or I-CreI restriction enzyme sites into an attP-site. These restriction enzyme sites allow for double-strand break-mediated removal of unwanted flanking transposon sequences, while leaving the desired genomic modifications or recombination cassettes. As a proof-of-principle, we mutated LRRK, tau, and sky by using different MiMIC elements. We replaced 6 kb of genomic DNA encompassing the tau locus and 35 kb encompassing the sky locus with a recombination cassette that permits easy integration of DNA at these loci and we also generated a functional LRRK(HA) knock in allele. Given that ~92% of the Drosophila genes are located within the vicinity (MiMIC element, our methodology enables the efficient manipulation of nearly every locus in the fruit fly genome without the need for inefficient donor-dependent homologous recombination events. Copyright © 2014 Vilain et al.

A new biotype of Fusarium oxysporum f. sp. lycopersici race 2 emerged by a transposon-driven mutation of avirulence gene AVR1.

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Kashiwa, Takeshi; Suzuki, Tatsuya; Sato, Akira; Akai, Kotaro; Teraoka, Tohru; Komatsu, Ken; Arie, Tsutomu

2016-07-01

Emergence of races in Fusarium oxysporum f. sp. lycopersici (Fol) is caused by loss or mutation of at least one avirulence (AVR) gene. The product of AVR1 is a small protein (Avr1) secreted by Fol in tomato xylem sap during infection. This protein triggers Fol race 1 specific resistance (I) in tomato, indicating that AVR1 is an AVR gene. Deletion of AVR1 in race 1 resulted in the emergence of race 2, and an additional mutation in AVR2 generated race 3. Previously, we reported a new biotype of race 3, KoChi-1, in which AVR1 was truncated by a transposon Hormin, which suggested a new route to evolution of races in Fol However, to date no race 2 isolate carrying Hormin-truncated AVR1 has been reported. In this report, we describe such isolates, represented by Chiba-5, in which Hormin insertion occurred in AVR1 at a position different from that in KoChi-1. AVR1 truncation in both isolates resulted in production of defective Avr1 proteins. Chiba-5 and KoChi-1 belong to different phylogenetic clades, A1 and A2, respectively, suggesting that insertion of Hormin in AVR1 in Chiba-5 and KoChi-1 occurred as independent evolutionary events. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Acetic acid production from marine algae. Progress report No. 1, July 1--September 30, 1977

Energy Technology Data Exchange (ETDEWEB)

Sanderson, J.E.; Augenstein, D.C.; Wise, D.L.

1977-10-14

Progress is reported in research designed to develop an economically competitive process for producing acetic acid from biomass for the purpose of sparing petroleum for other uses, to evaluate marine algae as a potential source of biomass, and to document the feasibility of running fermentations in fixed packed bed fermenters. It was demonstrated that marine algae can be fermented to acetic acid. Initial rates of up to 168 meq/1 day were observed. These rates are substantially in excess of the 47 meq/1 day used in the economic projections. Also, when using marine algae as a substrate, acid levels were generated equivalent to the highest reported with other substrates. It was also demonstrated that a 4-foot fixed packet bed fermenter may be operated with marine algae as a substrate at 20 percent solids or 200 meq/1.

Facile construction of a random protein domain insertion library using an engineered transposon.

Science.gov (United States)

Shah, Vandan; Pierre, Brennal; Kim, Jin Ryoun

2013-01-15

Insertional fusion between multiple protein domains represents a novel means of creating integrated functionalities. Currently, there is no robust guideline for selection of insertion sites ensuring the desired functional outcome of insertional fusion. Therefore, construction and testing of random domain insertion libraries, in which a host protein domain is randomly inserted into a guest protein domain, significantly benefit extensive exploration of sequence spaces for insertion sites. Short peptide residues are usually introduced between protein domains to alleviate structural conflicts, and the interdomain linker residues may affect the functional outcome of protein insertion complexes. Unfortunately, optimal control of interdomain linker residues is not always available in conventional methods used to construct random domain insertion libraries. Moreover, most conventional methods employ blunt-end rather than sticky-end ligation between host and guest DNA fragments, thus lowering library construction efficiency. Here, we report the facile construction of random domain insertion libraries using an engineered transposon. We show that random domain insertion with optimal control of interdomain linker residues was possible with our engineered transposon-based method. In addition, our method employs sticky-end rather than blunt-end ligation between host and guest DNA fragments, thus allowing for facile construction of relatively large sized libraries. Copyright © 2012 Elsevier Inc. All rights reserved.

Transposons and integrons in colistin-resistant clones of Klebsiella pneumoniae and Acinetobacter baumannii with epidemic or sporadic behaviour.

Science.gov (United States)

Arduino, Sonia M; Quiroga, María Paula; Ramírez, María Soledad; Merkier, Andrea Karina; Errecalde, Laura; Di Martino, Ana; Smayevsky, Jorgelina; Kaufman, Sara; Centrón, Daniela

2012-10-01

Multiple transposons, integrons and carbapenemases were found in Klebsiella pneumoniae colistin-resistant isolates as well as a genomic resistance island of the AbaR type in Acinetobacter baumannii colistin-resistant isolates from different hospitals from Buenos Aires City. PFGE analysis showed a polyclonal dissemination of antimicrobial resistance mechanisms among K. pneumoniae isolates, while in A. baumannii isolates the epidemic clone 1 from South America was found. Resistance determinants associated with horizontal gene transfer are contributing to the evolution to pandrug resistance in both epidemic and sporadic clones.

Comprehensive MALDI-TOF biotyping of the non-redundant Harvard Pseudomonas aeruginosa PA14 transposon insertion mutant library.

Science.gov (United States)

Oumeraci, Tonio; Jensen, Vanessa; Talbot, Steven R; Hofmann, Winfried; Kostrzewa, Markus; Schlegelberger, Brigitte; von Neuhoff, Nils; Häussler, Susanne

2015-01-01

Pseudomonas aeruginosa is a gram-negative bacterium that is ubiquitously present in the aerobic biosphere. As an antibiotic-resistant facultative pathogen, it is a major cause of hospital-acquired infections. Its rapid and accurate identification is crucial in clinical and therapeutic environments. In a large-scale MALDI-TOF mass spectrometry-based screen of the Harvard transposon insertion mutant library of P. aeruginosa strain PA14, intact-cell proteome profile spectra of 5547 PA14 transposon mutants exhibiting a plethora of different phenotypes were acquired and analyzed. Of all P. aeruginosa PA14 mutant profiles 99.7% were correctly identified as P. aeruginosa with the Biotyper software on the species level. On the strain level, 99.99% of the profiles were mapped to five different individual P. aeruginosa Biotyper database entries. A principal component analysis-based approach was used to determine the most important discriminatory mass features between these Biotyper groups. Although technical replicas were consistently categorized to specific Biotyper groups in 94.2% of the mutant profiles, biological replicas were not, indicating that the distinct proteotypes are affected by growth conditions. The PA14 mutant profile collection presented here constitutes the largest coherent P. aeruginosa MALDI-TOF spectral dataset publicly available today. Transposon insertions in thousands of different P. aeruginosa genes did not affect species identification from MALDI-TOF mass spectra, clearly demonstrating the robustness of the approach. However, the assignment of the individual spectra to sub-groups proved to be non-consistent in biological replicas, indicating that the differentiation between biotyper groups in this nosocomial pathogen is unassured.

Stable, Nonviral Expression of Mutated Tumor Neoantigen-specific T-cell Receptors Using the Sleeping Beauty Transposon/Transposase System

Science.gov (United States)

Deniger, Drew C; Pasetto, Anna; Tran, Eric; Parkhurst, Maria R; Cohen, Cyrille J; Robbins, Paul F; Cooper, Laurence JN; Rosenberg, Steven A

2016-01-01

Neoantigens unique to each patient's tumor can be recognized by autologous T cells through their T-cell receptor (TCR) but the low frequency and/or terminal differentiation of mutation-specific T cells in tumors can limit their utility as adoptive T-cell therapies. Transfer of TCR genes into younger T cells from peripheral blood with a high proliferative potential could obviate this problem. We generated a rapid, cost-effective strategy to genetically engineer cancer patient T cells with TCRs using the clinical Sleeping Beauty transposon/transposase system. Patient-specific TCRs reactive against HLA-A*0201-restriced neoantigens AHNAKS2580F or ERBB2H473Y or the HLA-DQB*0601-restricted neoantigen ERBB2IPE805G were assembled with murine constant chains and cloned into Sleeping Beauty transposons. Patient peripheral blood lymphocytes were coelectroporated with SB11 transposase and Sleeping Beauty transposon, and transposed T cells were enriched by sorting on murine TCRβ (mTCRβ) expression. Rapid expansion of mTCRβ+ T cells with irradiated allogeneic peripheral blood lymphocytes feeders, OKT3, interleukin-2 (IL-2), IL-15, and IL-21 resulted in a preponderance of effector (CD27−CD45RA−) and less-differentiated (CD27+CD45RA+) T cells. Transposed T cells specifically mounted a polyfunctional response against cognate mutated neoantigens and tumor cell lines. Thus, Sleeping Beauty transposition of mutation-specific TCRs can facilitate the use of personalized T-cell therapy targeting unique neoantigens. PMID:26945006

Drifter, a novel, low copy hAT-like transposon in Fusarium oxysporum is activated during starvation.

Science.gov (United States)

Rep, Martijn; van der Does, H Charlotte; Cornelissen, Ben J C

2005-06-01

The facultative pathogenic fungus Fusarium oxysporum is known to harbour many different transposable and/or repetitive elements. We have identified Drifter, a novel DNA transposon of the hAT family in F. oxysporum. It was found adjoining SIX1-H, a truncated homolog of the SIX1 avirulence gene in F. oxysporum f. sp. lycopersici. Absence of a target site duplication as well as the 5' part of SIX1-H suggests that transposition of Drifter into the ancestor of SIX1-H was followed by loss of a chromosomal segment through recombination between Drifters. F. oxysporum isolates belonging to various formae speciales harbour between 0 and 5 full-length copies of Drifter and/or one or more copies with an internal deletion. Transcription of Drifter is activated during starvation for carbon or nitrogen.

Discovery and widespread occurrence of polyhalogenated 1,1'-dimethyl-2,2'-bipyrroles (PDBPs) in marine biota

International Nuclear Information System (INIS)

Hauler, Carolin; Martin, René; Knölker, Hans-Joachim; Gaus, Caroline; Mueller, Jochen F.; Vetter, Walter

2013-01-01

Polyhalogenated 1,1′-dimethyl-2,2′-bipyrroles (PDBPs) are halogenated natural products (HNPs) previously shown to bioaccumulate in marine mammals and birds. Since their discovery in 1999, six hexahalogenated and a few lesser halogenated congeners have been identified in diverse marine mammal samples. Here we report the identification of 17 additional hexahalogenated PDBPs in the blubber extract of a humpback dolphin (Sousa chinensis) from Queensland, Australia. Thirteen of these new PDBPs were also detected in an Australian sea cucumber (Holothuria sp.). Additional samples were also tested positive on several new PDBPs, including an Australian venus tuskfish (Choerodon venustus) as well as a white whale (Delphinapterus leucas) and a sperm whale (Physeter macrocephalus) from the Northern Hemisphere. GC/ECNI-MS-SIM quantification of the molecular ions was carried out with the help of synthesized standards. The sum concentration of PDBPs was 1.1 mg/kg lipid in the humpback dolphin and 0.48 mg/kg lipid in the sea cucumber. -- Highlights: •Polyhalogenated 1,1′-dimethyl-2,2′-bipyrroles (PDBPs) are natural products. •17 New hexahalogenated PDBPs were identified in marine biota from Australia. •A humpback dolphin (Sousa chinensis) contained 1.1 mg/kg lipid PDBPs. •New PDBPs were also detected in marine mammals from the Northern Hemisphere. -- Detection of new polyhalogenated 1,1′-dimethyl-2,2′-bipyrroles indicates a higher toxic risk of these halogenated natural products in the marine environment than previously known

Gene Therapy with the Sleeping Beauty Transposon System.

Science.gov (United States)

Kebriaei, Partow; Izsvák, Zsuzsanna; Narayanavari, Suneel A; Singh, Harjeet; Ivics, Zoltán

2017-11-01

The widespread clinical implementation of gene therapy requires the ability to stably integrate genetic information through gene transfer vectors in a safe, effective, and economical manner. The latest generation of Sleeping Beauty (SB) transposon vectors fulfills these requirements, and may overcome limitations associated with viral gene transfer vectors and transient nonviral gene delivery approaches that are prevalent in ongoing clinical trials. The SB system enables high-level stable gene transfer and sustained transgene expression in multiple primary human somatic cell types, thereby representing a highly attractive gene transfer strategy for clinical use. Here, we review the most important aspects of using SB for gene therapy, including vectorization as well as genomic integration features. We also illustrate the path to successful clinical implementation by highlighting the application of chimeric antigen receptor (CAR)-modified T cells in cancer immunotherapy. Copyright © 2017 Elsevier Ltd. All rights reserved.

Transposon-mediated random gene disruption with moderate halophilic bacteria and its application for halophilic bacterial siderophore analysis.

Science.gov (United States)

Matsui, Toru; Nishino, Tomohiko

2016-12-01

Analytical conditions using chromo azurol S was validated for quantification of siderophore in aqueous samples, followed by the characterization of siderophore derived from newly isolated moderately halophilic bacteria. Conditions with good linearity between the absorbance and the siderophore concentration were obtained at a siderophore concentration less than 20 µM, in the wavelength range between 630 and 660 nm with developing time for at least 2 h. Of the halophilic bacteria isolated from Tunisian soil, Halomonas sp., namely strain 21a was selected as siderophore producing halophiles. The strain produced siderophore significantly in the absence of iron in minimal medium. Siderophore-deficient mutant, namely IIa10, of the strain 21a was obtained from gene disruptant library constructed using transposon complex by electroporation. Genomic sequence analysis of the mutant IIa10 revealed that the transposon-inserted gene was TonB-dependent receptor. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Enhanced Expression of Anti-CD19 Chimeric Antigen Receptor in piggyBac Transposon-Engineered T Cells

Directory of Open Access Journals (Sweden)

Daisuke Morita

2018-03-01

Full Text Available Adoptive T cell therapy using chimeric antigen receptor (CAR-modified T cells is a promising cancer immunotherapy. We previously developed a non-viral method of gene transfer into T cells using a piggyBac transposon system to improve the cost-effectiveness of CAR-T cell therapy. Here, we have further improved our technology by a novel culture strategy to increase the transfection efficiency and to reduce the time of T cell manufacturing. Using a CH2CH3-free CD19-specific CAR transposon vector and combining irradiated activated T cells (ATCs as feeder cells and virus-specific T cell receptor (TCR stimulation, we achieved 51.4% ± 14% CAR+ T cells and 2.8-fold expansion after 14 culture days. Expanded CD19.CAR-T cells maintained a significant fraction of CD45RA+CCR7+ T cells and demonstrated potent antitumor activity against CD19+ leukemic cells both in vitro and in vivo. Therefore, piggyBac-based gene transfer may provide an alternative to viral gene transfer for CAR-T cell therapy.

Prolonged Expression of Secreted Enzymes in Dogs After Liver-Directed Delivery of Sleeping Beauty Transposons: Implications for Non-Viral Gene Therapy of Systemic Disease.

Science.gov (United States)

Aronovich, Elena L; Hyland, Kendra A; Hall, Bryan C; Bell, Jason B; Olson, Erik R; Rusten, Myra Urness; Hunter, David W; Ellinwood, N Matthew; McIvor, R Scott; Hackett, Perry B

2017-07-01

The non-viral, integrating Sleeping Beauty (SB) transposon system is efficient in treating systemic monogenic disease in mice, including hemophilia A and B caused by deficiency of blood clotting factors and mucopolysaccharidosis types I and VII caused by α-L-iduronidase (IDUA) and β-glucuronidase (GUSB) deficiency, respectively. Modified approaches of the hydrodynamics-based procedure to deliver transposons to the liver in dogs were recently reported. Using the transgenic canine reporter secreted alkaline phosphatase (cSEAP), transgenic protein in the plasma was demonstrated for up to 6 weeks post infusion. This study reports that immunosuppression of dogs with gadolinium chloride (GdCl 3 ) prolonged the presence of cSEAP in the circulation up to 5.5 months after a single vector infusion. Transgene expression declined gradually but appeared to stabilize after about 2 months at approximately fourfold baseline level. Durability of transgenic protein expression in the plasma was inversely associated with transient increase of liver enzymes alanine transaminase and aspartate transaminase in response to the plasmid delivery procedure, which suggests a deleterious effect of hepatocellular toxicity on transgene expression. GdCl 3 treatment was ineffective for repeat vector infusions. In parallel studies, dogs were infused with potentially therapeutic transposons. Activities of transgenic IDUA and GUSB in plasma peaked at 50-350% of wildtype, but in the absence of immunosuppression lasted only a few days. Transposition was detectable by excision assay only when the most efficient transposase, SB100X, was used. Dogs infused with transposons encoding canine clotting factor IX (cFIX) were treated with GdCl 3 and showed expression profiles similar to those in cSEAP-infused dogs, with expression peaking at 40% wt (2 μg/mL). It is concluded that GdCl 3 can support extended transgene expression after hydrodynamic introduction of SB transposons in dogs, but that alternative

The BDGP gene disruption project: Single transposon insertions associated with 40 percent of Drosophila genes

Energy Technology Data Exchange (ETDEWEB)

Bellen, Hugo J.; Levis, Robert W.; Liao, Guochun; He, Yuchun; Carlson, Joseph W.; Tsang, Garson; Evans-Holm, Martha; Hiesinger, P. Robin; Schulze, Karen L.; Rubin, Gerald M.; Hoskins, Roger A.; Spradling, Allan C.

2004-01-13

The Berkeley Drosophila Genome Project (BDGP) strives to disrupt each Drosophila gene by the insertion of a single transposable element. As part of this effort, transposons in more than 30,000 fly strains were localized and analyzed relative to predicted Drosophila gene structures. Approximately 6,300 lines that maximize genomic coverage were selected to be sent to the Bloomington Stock Center for public distribution, bringing the size of the BDGP gene disruption collection to 7,140 lines. It now includes individual lines predicted to disrupt 5,362 of the 13,666 currently annotated Drosophila genes (39 percent). Other lines contain an insertion at least 2 kb from others in the collection and likely mutate additional incompletely annotated or uncharacterized genes and chromosomal regulatory elements. The remaining strains contain insertions likely to disrupt alternative gene promoters or to allow gene mis-expression. The expanded BDGP gene disruption collection provides a public resource that will facilitate the application of Drosophila genetics to diverse biological problems. Finally, the project reveals new insight into how transposons interact with a eukaryotic genome and helps define optimal strategies for using insertional mutagenesis as a genomic tool.

nanos-Driven expression of piggyBac transposase induces mobilization of a synthetic autonomous transposon in the malaria vector mosquito, Anopheles stephensi.

Science.gov (United States)

Macias, Vanessa M; Jimenez, Alyssa J; Burini-Kojin, Bianca; Pledger, David; Jasinskiene, Nijole; Phong, Celine Hien; Chu, Karen; Fazekas, Aniko; Martin, Kelcie; Marinotti, Osvaldo; James, Anthony A

2017-08-01

Transposons are a class of selfish DNA elements that can mobilize within a genome. If mobilization is accompanied by an increase in copy number (replicative transposition), the transposon may sweep through a population until it is fixed in all of its interbreeding members. This introgression has been proposed as the basis for drive systems to move genes with desirable phenotypes into target species. One such application would be to use them to move a gene conferring resistance to malaria parasites throughout a population of vector mosquitos. We assessed the feasibility of using the piggyBac transposon as a gene-drive mechanism to distribute anti-malarial transgenes in populations of the malaria vector, Anopheles stephensi. We designed synthetic gene constructs that express the piggyBac transposase in the female germline using the control DNA of the An. stephensi nanos orthologous gene linked to marker genes to monitor inheritance. Two remobilization events were observed with a frequency of one every 23 generations, a rate far below what would be useful to drive anti-pathogen transgenes into wild mosquito populations. We discuss the possibility of optimizing this system and the impetus to do so. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

A Tn5051-like mer-containing transposon identified in a heavy metal tolerant strain Achromobacter sp. AO22

Directory of Open Access Journals (Sweden)

Bhave Mrinal

2009-03-01

Full Text Available Abstract Background Achromobacter sp. AO22 (formerly Alcaligenes sp. AO22, a bacterial strain isolated from a lead-contaminated industrial site in Australia, was previously found to be resistant to moderate to high levels of mercury, copper and other heavy metals. However, the nature and location of the genetic basis for mercuric ion resistance in this strain, had not been previously identified. Findings Achromobacter sp. AO22 contains a functional mer operon with all four essential genes (merRTPA and shows >99% DNA sequence identity to that of Tn501. The mer operon was present on a transposon, designated TnAO22, captured by introducing a broad-host-range IncP plasmid into Achromobacter sp. AO22 and subsequently transferring it to E. coli recipients. The transposition frequency of TnAO22 was 10-2 to 10-3 per target plasmid transferred. Analysis of TnAO22 sequence revealed it belonged to the Tn21 subgroup of the Tn3 superfamily of transposons, with the transposition module having >99% identity with Tn5051 of a Pseudomonas putida strain isolated from a water sample in New York. Conclusion TnAO22 is thus a new variant of Tn5051 of the Tn3 superfamily and the transposon and its associated mercury resistance system are among the few such systems reported in a soil bacterium. Achromobacter sp. AO22 can thus be exploited for applications such as in situ mercury bioremediation of contaminated sites, or the mobile unit and mer operon could be mobilized to other bacteria for similar purposes.

Radioactivity in the Marine Environment. Chapter 1

International Nuclear Information System (INIS)

Zal U'yun Wan Mahmood; Abdul Kadir Ishak; Norfaizal Mohamad; Wo, Y.M.; Kamarudin Samuding

2015-01-01

Radionuclide (radioactive isotopes or radioisotopes is widely distributed on the ground primarily in marine environments. Nowadays, more than 340 isotopes has been identified exist in our earth especially in marine environment. From that total, 80 isotopes was radioactive. The existence of radioactivity in the marine environment is through the direct and indirect distribution of radionuclides

1st Advanced Marine Renewable Energy Instrumentation Experts Workshop: April 5-7, 2011

International Nuclear Information System (INIS)

2011-01-01

The U.S. marine energy industry is actively pursuing development of offshore wind and marine hydrokinetic (MHK) energy systems. Experience in the wind energy sector demonstrates that new technology development requires thorough measurement and characterization of the environmental conditions prevalent at installation sites and of technology operating in the field. Presently, there are no turn-key instrumentation system solutions that meet the measurement needs of the marine energy industry. The 1st Advanced Marine Renewable Energy Instrumentation Experts Workshop brought together technical experts from government laboratories, academia, and industry representatives from marine energy, wind, offshore oil and gas, and instrumentation developers to present and discuss the instrumentation needs of the marine energy industry. The goals of the meeting were to: (1) Share the latest relevant knowledge among technical experts; (2) Review relevant state-of-the-art field measurement technologies and methods; (3) Review lessons learned from recent field deployments; (4) Identify synergies across different industries; (5) Identify gaps between existing and needed instrumentation capabilities; (6) Understand who are the leading experts; (7) Provide a forum where stakeholders from the marine energy industry could provide substantive input in the development of new marine energy field deployable instrumentation packages.

1st Advanced Marine Renewable Energy Instrumentation Experts Workshop: April 5-7, 2011

Energy Technology Data Exchange (ETDEWEB)

2011-10-01

The U.S. marine energy industry is actively pursuing development of offshore wind and marine hydrokinetic (MHK) energy systems. Experience in the wind energy sector demonstrates that new technology development requires thorough measurement and characterization of the environmental conditions prevalent at installation sites and of technology operating in the field. Presently, there are no turn-key instrumentation system solutions that meet the measurement needs of the marine energy industry. The 1st Advanced Marine Renewable Energy Instrumentation Experts Workshop brought together technical experts from government laboratories, academia, and industry representatives from marine energy, wind, offshore oil and gas, and instrumentation developers to present and discuss the instrumentation needs of the marine energy industry. The goals of the meeting were to: (1) Share the latest relevant knowledge among technical experts; (2) Review relevant state-of-the-art field measurement technologies and methods; (3) Review lessons learned from recent field deployments; (4) Identify synergies across different industries; (5) Identify gaps between existing and needed instrumentation capabilities; (6) Understand who are the leading experts; (7) Provide a forum where stakeholders from the marine energy industry could provide substantive input in the development of new marine energy field deployable instrumentation packages.

The Drosophila Su(var)3-7 gene is required for oogenesis and female fertility, genetically interacts with piwi and aubergine, but impacts only weakly transposon silencing.

Science.gov (United States)

Basquin, Denis; Spierer, Anne; Begeot, Flora; Koryakov, Dmitry E; Todeschini, Anne-Laure; Ronsseray, Stéphane; Vieira, Cristina; Spierer, Pierre; Delattre, Marion

2014-01-01

Heterochromatin is made of repetitive sequences, mainly transposable elements (TEs), the regulation of which is critical for genome stability. We have analyzed the role of the heterochromatin-associated Su(var)3-7 protein in Drosophila ovaries. We present evidences that Su(var)3-7 is required for correct oogenesis and female fertility. It accumulates in heterochromatic domains of ovarian germline and somatic cells nuclei, where it co-localizes with HP1. Homozygous mutant females display ovaries with frequent degenerating egg-chambers. Absence of Su(var)3-7 in embryos leads to defects in meiosis and first mitotic divisions due to chromatin fragmentation or chromosome loss, showing that Su(var)3-7 is required for genome integrity. Females homozygous for Su(var)3-7 mutations strongly impair repression of P-transposable element induced gonadal dysgenesis but have minor effects on other TEs. Su(var)3-7 mutations reduce piRNA cluster transcription and slightly impact ovarian piRNA production. However, this modest piRNA reduction does not correlate with transposon de-silencing, suggesting that the moderate effect of Su(var)3-7 on some TE repression is not linked to piRNA production. Strikingly, Su(var)3-7 genetically interacts with the piwi and aubergine genes, key components of the piRNA pathway, by strongly impacting female fertility without impairing transposon silencing. These results lead us to propose that the interaction between Su(var)3-7 and piwi or aubergine controls important developmental processes independently of transposon silencing.

Toward forward genetic screens in malaria-causing parasites using the piggyBac transposon

Directory of Open Access Journals (Sweden)

de Koning-Ward Tania F

2011-03-01

Full Text Available Abstract The ability to analyze gene function in malaria-causing Plasmodium parasites has received a boost with a recent paper in BMC Genomics that describes a genome-wide mutagenesis system in the rodent malaria species Plasmodium berghei using the transposon piggyBac. This advance holds promise for identifying and validating new targets for intervention against malaria. But further improvements are still needed for the full power of genome-wide molecular genetic screens to be utilized in this organism. See research article: http://www.biomedcentral.com/1471-2164/12/155

Genetic structure of landraces in foxtail millet (Setaria italica (L.) P. Beauv.) revealed with transposon display and interpretation to crop evolution of foxtail millet.

Science.gov (United States)

Hirano, Ryoko; Naito, Ken; Fukunaga, Kenji; Watanabe, Kazuo N; Ohsawa, Ryo; Kawase, Makoto

2011-06-01

Although the origin and domestication process of foxtail millet (Setaria italica subsp. italica (L.) P. Beauv.) has been studied by several groups, the issue is still ambiguous. It is essential to resolve this issue by studying a large number of accessions with sufficient markers covering the entire genome. Genetic structures were analyzed by transposon display (TD) using 425 accessions of foxtail millet and 12 of the wild ancestor green foxtail (Setaria italica subsp. viridis (L.) P. Beauv.). We used three recently active transposons (TSI-1, TSI-7, and TSI-10) as genome-wide markers and succeeded in demonstrating geographical structures of the foxtail millet. A neighbor-joining dendrogram based on TD grouped the foxtail millet accessions into eight major clusters, each of which consisted of accessions collected from adjacent geographical areas. Eleven out of 12 green foxtail accessions were grouped separately from the clusters of foxtail millet. These results indicated strong regional differentiations and a long history of cultivation in each region. Furthermore, we discuss the relationship between foxtail millet and green foxtail and suggest a monophyletic origin of foxtail millet domestication.

An active ac/ds transposon system for activation tagging in tomato cultivar m82 using clonal propagation.

Science.gov (United States)

Carter, Jared D; Pereira, Andy; Dickerman, Allan W; Veilleux, Richard E

2013-05-01

Tomato (Solanum lycopersicum) is a model organism for Solanaceae in both molecular and agronomic research. This project utilized Agrobacterium tumefaciens transformation and the transposon-tagging construct Activator (Ac)/Dissociator (Ds)-ATag-Bar_gosGFP to produce activation-tagged and knockout mutants in the processing tomato cultivar M82. The construct carried hygromycin resistance (hyg), green fluorescent protein (GFP), and the transposase (TPase) of maize (Zea mays) Activator major transcript X054214.1 on the stable Ac element, along with a 35S enhancer tetramer and glufosinate herbicide resistance (BAR) on the mobile Ds-ATag element. An in vitro propagation strategy was used to produce a population of 25 T0 plants from a single transformed plant regenerated in tissue culture. A T1 population of 11,000 selfed and cv M82 backcrossed progeny was produced from the functional T0 line. This population was screened using glufosinate herbicide, hygromycin leaf painting, and multiplex polymerase chain reaction (PCR). Insertion sites of transposed Ds-ATag elements were identified through thermal asymmetric interlaced PCR, and resulting product sequences were aligned to the recently published tomato genome. A population of 509 independent, Ds-only transposant lines spanning all 12 tomato chromosomes has been developed. Insertion site analysis demonstrated that more than 80% of these lines harbored Ds insertions conducive to activation tagging. The capacity of the Ds-ATag element to alter transcription was verified by quantitative real-time reverse transcription-PCR in two mutant lines. The transposon-tagged lines have been immortalized in seed stocks and can be accessed through an online database, providing a unique resource for tomato breeding and analysis of gene function in the background of a commercial tomato cultivar.

Marine environment news. Vol. 2, no. 1

International Nuclear Information System (INIS)

2004-03-01

In this issue of the IAEA's Marine Environment Newsletter topics including radiotracers as new barometers of ocean-climate coupling, bio-indicatos species in detecting marine radioactvity and pollution as well as training activities are covered

Increased encapsulated cell biodelivery of nerve growth factor in the brain by transposon-mediated gene transfer

DEFF Research Database (Denmark)

Fjord-Larsen, L; Kusk, Poul Henrik; Emerich, D F

2012-01-01

transposon expression technology to establish a new clinical grade cell line, NGC0211, with at least 10 times higher NGF production than that of NGC-0295. To test whether encapsulation of this cell line provides a relevant dose escalation step in delivering NGF for treatment of the cognitive decline in AD...... cases correlate with highly improved potency.Gene Therapy advance online publication, 24 November 2011; doi:10.1038/gt.2011.178....

Transgenerational changes in plant physiology and in transposon expression in response to UV-C stress in Arabidopsis thaliana.

Science.gov (United States)

Migicovsky, Zoe; Kovalchuk, Igor

2014-01-01

Stress has a negative impact on crop yield by altering a gain in biomass and affecting seed set. Recent reports suggest that exposure to stress also influences the response of the progeny. In this paper, we analyzed seed size, leaf size, bolting time and transposon expression in 2 consecutive generations of Arabidopsis thaliana plants exposed to moderate UV-C stress. Since previous reports suggested a potential role of Dicer-like (DCL) proteins in the establishment of transgenerational response, we used dcl2, dcl3 and dcl4 mutants in parallel with wild-type plants. These studies revealed that leaf number decreased in the progeny of UV-C stressed plants, and bolting occurred later. Transposons were also re-activated in the progeny of stressed plants. Changes in the dcl mutants were less prominent than in wild-type plants. DCL2 and DCL3 appeared to be more important in the transgenerational stress memory than DCL4 because transgenerational changes were less profound in the dcl2 and dcl3 mutants.

Two hAT transposon genes were transferred from Brassicaceae to broomrapes and are actively expressed in some recipients

Science.gov (United States)

Sun, Ting; Renner, Susanne S.; Xu, Yuxing; Qin, Yan; Wu, Jianqiang; Sun, Guiling

2016-01-01

A growing body of evidence is pointing to an important role of horizontal gene transfer (HGT) in the evolution of higher plants. However, reports of HGTs of transposable elements (TEs) in plants are still scarce, and only one case is known of a class II transposon horizontally transferred between grasses. To investigate possible TE transfers in dicots, we performed transcriptome screening in the obligate root parasite Phelipanche aegyptiaca (Orobanchaceae), data-mining in the draft genome assemblies of four other Orobanchaceae, gene cloning, gene annotation in species with genomic information, and a molecular phylogenetic analysis. We discovered that the broomrape genera Phelipanche and Orobanche acquired two related nuclear genes (christened BO transposase genes), a new group of the hAT superfamily of class II transposons, from Asian Sisymbrieae or a closely related tribe of Brassicaceae, by HGT. The collinearity of the flanking genes, lack of a classic border structure, and low expression levels suggest that BO transposase genes cannot transpose in Brassicaceae, whereas they are highly expressed in P. aegyptiaca. PMID:27452947

Cut-and-Paste Transposons in Fungi with Diverse Lifestyles.

Science.gov (United States)

Muszewska, Anna; Steczkiewicz, Kamil; Stepniewska-Dziubinska, Marta; Ginalski, Krzysztof

2017-12-01

Transposable elements (TEs) shape genomes via recombination and transposition, lead to chromosomal rearrangements, create new gene neighborhoods, and alter gene expression. They play key roles in adaptation either to symbiosis in Amanita genus or to pathogenicity in Pyrenophora tritici-repentis. Despite growing evidence of their importance, the abundance and distribution of mobile elements replicating in a "cut-and-paste" fashion is barely described so far. In order to improve our knowledge on this old and ubiquitous class of transposable elements, 1,730 fungal genomes were scanned using both de novo and homology-based approaches. DNA TEs have been identified across the whole data set and display uneven distribution from both DNA TE classification and fungal taxonomy perspectives. DNA TE content correlates with genome size, which confirms that many transposon families proliferate simultaneously. In contrast, it is independent from intron density, average gene distance and GC content. TE count is associated with species' lifestyle and tends to be elevated in plant symbionts and decreased in animal parasites. Lastly, we found that fungi with both RIP and RNAi systems have more total DNA TE sequences but less elements retaining a functional transposase, what reflects stringent control over transposition. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

Free-living Marine Nematodes. Part 1 British Enoplids

African Journals Online (AJOL)

This is the first of three volumes dealing with the most abundant group of animals on the sea-bed and sea-shore, the free-living marine nematodes, and is devoted to those marine nematodes belonging to the subclass Enoplia. Separate volumes will deal with the orders Chromadorida and. Monhysterida. To most marine ...

The Foldback-like element Galileo belongs to the P superfamily of DNA transposons and is widespread within the Drosophila genus.

Science.gov (United States)

Marzo, Mar; Puig, Marta; Ruiz, Alfredo

2008-02-26

Galileo is the only transposable element (TE) known to have generated natural chromosomal inversions in the genus Drosophila. It was discovered in Drosophila buzzatii and classified as a Foldback-like element because of its long, internally repetitive, terminal inverted repeats (TIRs) and lack of coding capacity. Here, we characterized a seemingly complete copy of Galileo from the D. buzzatii genome. It is 5,406 bp long, possesses 1,229-bp TIRs, and encodes a 912-aa transposase similar to those of the Drosophila melanogaster 1360 (Hoppel) and P elements. We also searched the recently available genome sequences of 12 Drosophila species for elements similar to Dbuz\\Galileo by using bioinformatic tools. Galileo was found in six species (ananassae, willistoni, peudoobscura, persimilis, virilis, and mojavensis) from the two main lineages within the Drosophila genus. Our observations place Galileo within the P superfamily of cut-and-paste transposons and extend considerably its phylogenetic distribution. The interspecific distribution of Galileo indicates an ancient presence in the genus, but the phylogenetic tree built with the transposase amino acid sequences contrasts significantly with that of the species, indicating lineage sorting and/or horizontal transfer events. Our results also suggest that Foldback-like elements such as Galileo may evolve from DNA-based transposon ancestors by loss of the transposase gene and disproportionate elongation of TIRs.

1-hydroxypyrene as a biomarker of PAH exposure in the marine invertebrates N. diversicolor

DEFF Research Database (Denmark)

Tairova, Zhanna; Giessing, Anders; Hansen, Rikke

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous, persistent and toxic contaminants in the marine environment. Uptake of PAHs by marine deposit-feeding invertebrates can be determined by screening for PAH-derived metabolites. Methods for detection and quantification of PAH metabolites may...... invertebrates is on the other hand practically non-existing. The present study investigated formation of pyrene metabolites in the marine polychaete Nereis diversicolor, with special attention given to the detection of 1-hydroxypyrene. 1-hydroxypyrene was identified in tissues of Nereis diversicolor exposed...... this context the usefulness of SFS for identification and quantification was explored. Considering the complex composition of environmental PAH pollution, before the 1-hydoxypyrene in tissues of exposed marine invertebrates can be regarded as a suitable biomarker for assessing total PAH exposure...

The Drosophila Su(var)3–7 Gene Is Required for Oogenesis and Female Fertility, Genetically Interacts with piwi and aubergine, but Impacts Only Weakly Transposon Silencing

Science.gov (United States)

Begeot, Flora; Koryakov, Dmitry E.; Todeschini, Anne-Laure; Ronsseray, Stéphane; Vieira, Cristina; Spierer, Pierre; Delattre, Marion

2014-01-01

Heterochromatin is made of repetitive sequences, mainly transposable elements (TEs), the regulation of which is critical for genome stability. We have analyzed the role of the heterochromatin-associated Su(var)3–7 protein in Drosophila ovaries. We present evidences that Su(var)3–7 is required for correct oogenesis and female fertility. It accumulates in heterochromatic domains of ovarian germline and somatic cells nuclei, where it co-localizes with HP1. Homozygous mutant females display ovaries with frequent degenerating egg-chambers. Absence of Su(var)3–7 in embryos leads to defects in meiosis and first mitotic divisions due to chromatin fragmentation or chromosome loss, showing that Su(var)3–7 is required for genome integrity. Females homozygous for Su(var)3–7 mutations strongly impair repression of P-transposable element induced gonadal dysgenesis but have minor effects on other TEs. Su(var)3–7 mutations reduce piRNA cluster transcription and slightly impact ovarian piRNA production. However, this modest piRNA reduction does not correlate with transposon de-silencing, suggesting that the moderate effect of Su(var)3–7 on some TE repression is not linked to piRNA production. Strikingly, Su(var)3–7 genetically interacts with the piwi and aubergine genes, key components of the piRNA pathway, by strongly impacting female fertility without impairing transposon silencing. These results lead us to propose that the interaction between Su(var)3–7 and piwi or aubergine controls important developmental processes independently of transposon silencing. PMID:24820312

26 CFR 1.821-1 - Tax on mutual insurance companies other than life or marine or fire insurance companies subject...

Science.gov (United States)

2010-04-01

... life or marine or fire insurance companies subject to the tax imposed by section 831. 1.821-1 Section 1... and Other Than Fire Or Flood Insurance Companies Which Operate on Basis of Perpetual Policies Or Premium Deposits) § 1.821-1 Tax on mutual insurance companies other than life or marine or fire insurance...

46 CFR 167.60-1 - Issuance by Officer in Charge, Marine Inspection.

Science.gov (United States)

2010-10-01

... SCHOOLS PUBLIC NAUTICAL SCHOOL SHIPS Certificates of Inspection § 167.60-1 Issuance by Officer in Charge, Marine Inspection. (a) Every nautical school ship shall be inspected annually and if in the opinion of the Officer in Charge, Marine Inspection, the nautical school ship can be operated safely, he shall...

Transposon-mediated BAC transgenesis in human ES cells

Science.gov (United States)

Rostovskaya, Maria; Fu, Jun; Obst, Mandy; Baer, Isabell; Weidlich, Stefanie; Wang, Hailong; Smith, Andrew J. H.; Anastassiadis, Konstantinos; Stewart, A. Francis

2012-01-01

Transgenesis is a cornerstone of molecular biology. The ability to integrate a specifically engineered piece of DNA into the genome of a living system is fundamental to our efforts to understand life and exploit its implications for medicine, nanotechnology and bioprospecting. However, transgenesis has been hampered by position effects and multi-copy integration problems, which are mainly due to the use of small, plasmid-based transgenes. Large transgenes based on native genomic regions cloned into bacterial artificial chromosomes (BACs) circumvent these problems but are prone to fragmentation. Herein, we report that contrary to widely held notions, large BAC-sized constructs do not prohibit transposition. We also report the first reliable method for BAC transgenesis in human embryonic stem cells (hESCs). The PiggyBac or Sleeping Beauty transposon inverted repeats were integrated into BAC vectors by recombineering, followed by co-lipofection with the corresponding transposase in hESCs to generate robust fluorescent protein reporter lines for OCT4, NANOG, GATA4 and PAX6. BAC transposition delivers several advantages, including increased frequencies of single-copy, full-length integration, which will be useful in all transgenic systems but especially in difficult venues like hESCs. PMID:22753106

R-prime site-directed transposon Tn7 mutagenesis of the photosynthetic apparatus in Rhodopseudomonas capsulata

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Youvan, D C [Univ. of California, Berkeley; Elder, J T; Sandlin, D E; Zsebo, K; Alder, D P; Panopoulos, N J; Marrs, B L; Hearst, J E

1982-01-01

Site-directed mutagenesis of the photosynthetic apparatus (PSA) genes in Rhodopseudomonas capsulata is presented utilizing a transposon Tn7 mutagenized R-prime. The R-prime, pRPS404, bears most of the genes necessary for the differentiation of the photosynthetic apparatus. Mutagenesis of the R-prime with Tn7 in Escherichia coli, conjugation into R. capsulata, and homologous recombination with the wild-type alleles efficiently generates photosynthetic apparatus lesions. Wild-type alleles are lost spontaneously and the Tn7-induced lesions are revealed by subsequent intramolecular recombination between IS21 insertion elements that bracket the prime sequences in direct repeat. The molecular nature of the intermediates involved in the transposition, recombination and deletion have been investigated by Southern hybridization analysis. The spontaneous loss of wild-type alleles after homologous recombination with the chromosome may be of general use to other prokaryotic site-directed transposon mutagenesis schemes. The IS21-mediated deletion of the prime DNA is dependent on the RecA protein in E. coli, generating the parental R-factor bearing one IS21 element. A genetic-physical map exists for a portion of the prime photosynthetic apparatus DNA. When Tn7 is inserted into a bacteriochlorophyll gene in the R-prime and then crossed into R. capsulata, mutants are produced that accumulate a bacteriochlorophyll precursor, which is in excellent agreement with the existing genetic-physical map. This corroborates the mutagenesis scheme.

Genetic Transformation of Hordeum vulgare ssp. spontaneum for the Development of a Transposon-Based Insertional Mutagenesis System.

Science.gov (United States)

Cardinal, Marie-Josée; Kaur, Rajvinder; Singh, Jaswinder

2016-10-01

Domestication and intensive selective breeding of plants has triggered erosion of genetic diversity of important stress-related alleles. Researchers highlight the potential of using wild accessions as a gene source for improvement of cereals such as barley, which has major economic and social importance worldwide. Previously, we have successfully introduced the maize Ac/Ds transposon system for gene identification in cultivated barley. The objective of current research was to investigate the response of Hordeum vulgare ssp. spontaneum wild barley accessions in tissue culture to standardize parameters for introduction of Ac/Ds transposons through genetic transformation. We investigated the response of ten wild barley genotypes for callus induction, regenerative green callus induction and regeneration of fertile plants. The activity of exogenous Ac/Ds elements was observed through a transient assay on immature wild barley embryos/callus whereby transformed embryos/calli were identified by the expression of GUS. Transient Ds expression bombardment experiments were performed on 352 pieces of callus (3-5 mm each) or immature embryos in 4 genotypes of wild barley. The transformation frequency of putative transgenic callus lines based on transient GUS expression ranged between 72 and100 % in wild barley genotypes. This is the first report of a transformation system in H. vulgare ssp. spontaneum.

High-Level Heat Resistance of Spores of Bacillus amyloliquefaciens and Bacillus licheniformis Results from the Presence of a spoVA Operon in a Tn1546 Transposon

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Berendsen, Erwin M.; Koning, Rosella A.; Boekhorst, Jos; de Jong, Anne; Kuipers, Oscar P.; Wells-Bennik, Marjon H. J.

2016-01-01

Bacterial endospore formers can produce spores that are resistant to many food processing conditions, including heat. Some spores may survive heating processes aimed at production of commercially sterile foods. Recently, it was shown that a spoVA operon, designated spoVA2mob, present on a Tn1546 transposon in Bacillus subtilis, leads to profoundly increased wet heat resistance of B. subtilis spores. Such Tn1546 transposon elements including the spoVA2mob operon were also found in several strains of Bacillus amyloliquefaciens and Bacillus licheniformis, and these strains were shown to produce spores with significantly higher resistances to wet heat than their counterparts lacking this transposon. In this study, the locations and compositions of Tn1546 transposons encompassing the spoVA2mob operons in B. amyloliquefaciens and B. licheniformis were analyzed. Introduction of these spoVA2mob operons into B. subtilis 168 (producing spores that are not highly heat resistant) rendered mutant 168 strains that produced high-level heat resistant spores, demonstrating that these elements in B. amyloliquefaciens and B. licheniformis are responsible for high level heat resistance of spores. Assessment of growth of the nine strains of each species between 5.2°C and 57.7°C showed some differences between strains, especially at lower temperatures, but all strains were able to grow at 57.7°C. Strains of B. amyloliquefaciens and B. licheniformis that contain the Tn1546 elements (and produce high-level heat resistant spores) grew at temperatures similar to those of their Tn1546-negative counterparts that produce low-level heat resistant spores. The findings presented in this study allow for detection of B. amyloliquefaciens and B. licheniformis strains that produce highly heat resistant spores in the food chain. PMID:27994575

Global distribution and climate forcing of marine organic aerosol: 1. Model improvements and evaluation

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N. Meskhidze

2011-11-01

Full Text Available Marine organic aerosol emissions have been implemented and evaluated within the National Center of Atmospheric Research (NCAR's Community Atmosphere Model (CAM5 with the Pacific Northwest National Laboratory's 7-mode Modal Aerosol Module (MAM-7. Emissions of marine primary organic aerosols (POA, phytoplankton-produced isoprene- and monoterpenes-derived secondary organic aerosols (SOA and methane sulfonate (MS⁻ are shown to affect surface concentrations of organic aerosols in remote marine regions. Global emissions of submicron marine POA is estimated to be 7.9 and 9.4 Tg yr⁻¹, for the Gantt et al. (2011 and Vignati et al. (2010 emission parameterizations, respectively. Marine sources of SOA and particulate MS⁻ (containing both sulfur and carbon atoms contribute an additional 0.2 and 5.1 Tg yr⁻¹, respectively. Widespread areas over productive waters of the Northern Atlantic, Northern Pacific, and the Southern Ocean show marine-source submicron organic aerosol surface concentrations of 100 ng m⁻³, with values up to 400 ng m⁻³ over biologically productive areas. Comparison of long-term surface observations of water insoluble organic matter (WIOM with POA concentrations from the two emission parameterizations shows that despite revealed discrepancies (often more than a factor of 2, both Gantt et al. (2011 and Vignati et al. (2010 formulations are able to capture the magnitude of marine organic aerosol concentrations, with the Gantt et al. (2011 parameterization attaining better seasonality. Model simulations show that the mixing state of the marine POA can impact the surface number concentration of cloud condensation nuclei (CCN. The largest increases (up to 20% in CCN (at a supersaturation (S of 0.2% number concentration are obtained over biologically productive ocean waters when marine organic aerosol is assumed to be externally mixed with sea-salt. Assuming

Development of a high-frequency in vivo transposon mutagenesis system for Synechocystis sp. PCC 6803 and Synechococcus elongatus PCC 7942.

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Watabe, Kazuyuki; Mimuro, Mamoru; Tsuchiya, Tohru

2014-11-01

Synechocystis sp. PCC 6803 (Synechocystis) is the first sequenced photosynthetic organism and has two advantages: natural transformation and light-activated heterotrophic growth. Such characteristics have mainly promoted reverse genetic analysis in this organism, however, to date approximately 50% of genes are still annotated as 'unknown protein' or 'hypothetical protein'. Therefore, forward genetic analysis is required for the identification of significant genes responsible for photosynthesis and other physiological phenomena among the genes of unknown function. The in vivo transposon mutagenesis system is one of the major methods for random mutagenesis. However, present in vivo transposon mutagenesis systems for cyanobacteria face problems such as relatively low frequency of transposition and repeated transposition in the host cells. In this study, we constructed vectors based on a mini-Tn5-derived vector that was designed to prevent repeated transposition. Our vectors carry a hyperactive transposase and optimized recognition sequence of transposase, which were reported to enhance frequency of transposition. Using the vector, we succeeded in highly frequent transposition (9×10(-3) per recipient cell) in Synechocystis. Transposon insertion sites of 10 randomly selected mutants indicated that the insertion sites spread throughout the genome with low sequence dependency. Furthermore, one of the 10 mutants exhibited the slow-growing phenotype, and the mutant was functionally complemented by using our expression vector. Our system also worked with another model cyanobacterium, Synechococcus elongatus PCC 7942, with high frequency. These results indicate that the developed system can be applied to the forward genetic analysis of a broad range of cyanobacteria. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Physiological and behavioural impacts of Pacific ciguatoxin-1 (P-CTX-1) on marine medaka (Oryzias melastigma).

Science.gov (United States)

Mak, Yim Ling; Li, Jing; Liu, Chih-Ning; Cheng, Shuk Han; Lam, Paul K S; Cheng, Jinping; Chan, Leo L

2017-01-05

Ciguatoxins (CTXs) are natural biotoxins produced by benthic dinoflagellates of the genus Gambierdiscus, which are bioaccumulated and biotransformed along food chains in coral ecosystems. They are neurotoxins that activate voltage-gated sodium channels and disrupt ion conductance in the excitable tissues. Pacific ciguatoxin-1 (P-CTX-1) is the most prevalent and potent CTX congener present in fishes from the Pacific Ocean. In this study, P-CTX-1 was administrated to larval marine medaka (2h post-hatch) via microinjection. Exposure to P-CTX-1 at sub-ppb levels led to adverse behavioural changes, altered physiological performances and reduced survivability of the larval marine medaka as early as 24h after exposure. P-CTX-1 decreased the rate of heartbeat and locomotion of the exposed larvae, probably owing to a series of physiological processes and morphological changes such as pericardial oedema, failure of swim bladder inflation and spinal curvature. The exposed larval marine medaka also demonstrated reduced, delayed and paralyzed responses to external stimulations. This may render them more susceptible to predation. P-CTX-1 could be effectively distributed from the yolk sac to all parts of the fish body, including head and trunk, 24h after exposure. Repeated low-dose P-CTX-1 exposure resulted in larval mortality comparable to that of a single high-dose exposure. Copyright © 2016 Elsevier B.V. All rights reserved.

A New Class of SINEs with snRNA Gene-Derived Heads.

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Kojima, Kenji K

2015-05-27

Eukaryotic genomes are colonized by various transposons including short interspersed elements (SINEs). The 5' region (head) of the majority of SINEs is derived from one of the three types of RNA genes--7SL RNA, transfer RNA (tRNA), or 5S ribosomal RNA (rRNA)--and the internal promoter inside the head promotes the transcription of the entire SINEs. Here I report a new group of SINEs whose heads originate from either the U1 or U2 small nuclear RNA gene. These SINEs, named SINEU, are distributed among crocodilians and classified into three families. The structures of the SINEU-1 subfamilies indicate the recurrent addition of a U1- or U2-derived sequence onto the 5' end of SINEU-1 elements. SINEU-1 and SINEU-3 are ancient and shared among alligators, crocodiles, and gharials, while SINEU-2 is absent in the alligator genome. SINEU-2 is the only SINE family that was active after the split of crocodiles and gharials. All SINEU families, especially SINEU-3, are preferentially inserted into a family of Mariner DNA transposon, Mariner-N4_AMi. A group of Tx1 non-long terminal repeat retrotransposons designated Tx1-Mar also show target preference for Mariner-N4_AMi, indicating that SINEU was mobilized by Tx1-Mar. © The Author(s) 2015. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

A novel, simple, high-throughput method for isolation of genome-wide transposon insertion mutants of Escherichia coli K-12.

Science.gov (United States)

Miki, Takeyoshi; Yamamoto, Yoshihiro; Matsuda, Hideo

2008-01-01

We developed a novel, simple, high-throughput method for isolation of genome-wide transposon insertion mutants of Escherichia coli K-12. The basic idea of the method is to randomly disrupt the genes on the DNA fragments cloned on the Kohara library by inserting a mini-transposon first, and then transfer the disrupted genes from the lambda vector to the E. coli chromosome by homologous recombination. Using this method, we constructed a set of 8402 Km(r) cis-diploid mutants harboring a mini-Tn10 insertion mutation and the corresponding wild-type gene on a chromosome, as well as a set of 6954 haploid mutants derived from the cis-diploid mutants. The major advantage of the strategy used is that the indispensable genes or sites for growth can be identified. Preliminary results suggest that 415 open reading frames are indispensable for growth in E. coli cells. A total of 6404 haploid mutants were deposited to Genetic Strains Research Center, National Institute of Genetics, Japan (Chapter 26) and are available for public distribution upon request (http://shigen.lab.nig.ac.jp/ecoli/strain/nbrp/resource.jsp).

Linear and exponential TAIL-PCR: a method for efficient and quick amplification of flanking sequences adjacent to Tn5 transposon insertion sites.

Science.gov (United States)

Jia, Xianbo; Lin, Xinjian; Chen, Jichen

2017-11-02

Current genome walking methods are very time consuming, and many produce non-specific amplification products. To amplify the flanking sequences that are adjacent to Tn5 transposon insertion sites in Serratia marcescens FZSF02, we developed a genome walking method based on TAIL-PCR. This PCR method added a 20-cycle linear amplification step before the exponential amplification step to increase the concentration of the target sequences. Products of the linear amplification and the exponential amplification were diluted 100-fold to decrease the concentration of the templates that cause non-specific amplification. Fast DNA polymerase with a high extension speed was used in this method, and an amplification program was used to rapidly amplify long specific sequences. With this linear and exponential TAIL-PCR (LETAIL-PCR), we successfully obtained products larger than 2 kb from Tn5 transposon insertion mutant strains within 3 h. This method can be widely used in genome walking studies to amplify unknown sequences that are adjacent to known sequences.

Transcriptome-wide Identification and Expression Analysis of Brachypodium distachyon Transposons in Response to Viral Infection

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Tuğba Gürkök

2017-10-01

Full Text Available Transposable elements (TEs are the most abundant group of genomic elements in plants that can be found in genic or intergenic regions of their host genomes. Several stimuli such as biotic or abiotic stress have roles in either activating their transcription or transposition. Here the effect of the Panicum mosaic virus (PMV and its satellite virus (SPMV infection on the transposon transcription of the Brachypodium distachyon model plant was investigated. To evaluate the transcription activity of TEs, transcriptomic data of mock and virus inoculated plants were compared. Our results indicate that major components of TEs are retroelements in all RNA-seq libraries. The number of transcribed TEs detected in mock inoculated plants is higher than virus inoculated plants. In comparison with mock inoculated plants 13% of the TEs showed at least two folds alteration upon PMV infection and 21% upon PMV+SPMV infection. Rather than inoculation with PMV alone inoculation with PMV+SPMV together also increased various TE encoding transcripts expressions. MuDR-N78C_OS encoding transcript was strongly up-regulated against both PMV and PMV+SPMV infection. The synergism generated by PMV and SPMV together enhanced TE transcripts expressions than PMV alone. It was observed that viral infection induced the transcriptional activity of several transposons. The results suggest that increased expressions of TEs might have a role in response to biotic stress in B. distachyon. Identification of TEs which are taking part in stress can serve useful information for functional genomics and designing novel breeding strategies in developing stress resistance crops.

Proteus genomic island 1 (PGI1), a new resistance genomic island from two Proteus mirabilis French clinical isolates.

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Siebor, Eliane; Neuwirth, Catherine

2014-12-01

To analyse the genetic environment of the antibiotic resistance genes in two clinical Proteus mirabilis isolates resistant to multiple antibiotics. PCR, gene walking and whole-genome sequencing were used to determine the sequence of the resistance regions, the surrounding genetic structure and the flanking chromosomal regions. A genomic island of 81.1 kb named Proteus genomic island 1 (PGI1) located at the 3'-end of trmE (formerly known as thdF) was characterized. The large MDR region of PGI1 (55.4 kb) included a class 1 integron (aadB and aadA2) and regions deriving from several transposons: Tn2 (blaTEM-135), Tn21, Tn6020-like transposon (aphA1b), a hybrid Tn502/Tn5053 transposon, Tn501, a hybrid Tn1696/Tn1721 transposon [tetA(A)] carrying a class 1 integron (aadA1) and Tn5393 (strA and strB). Several ISs were also present (IS4321, IS1R and IS26). The PGI1 backbone (25.7 kb) was identical to that identified in Salmonella Heidelberg SL476 and shared some identity with the Salmonella genomic island 1 (SGI1) backbone. An IS26-mediated recombination event caused the division of the MDR region into two parts separated by a large chromosomal DNA fragment of 197 kb, the right end of PGI1 and this chromosomal sequence being in inverse orientation. PGI1 is a new resistance genomic island from P. mirabilis belonging to the same island family as SGI1. The role of PGI1 in the spread of antimicrobial resistance genes among Enterobacteriaceae of medical importance needs to be evaluated. © The Author 2014. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Transposon defense by endo-siRNAs, piRNAs and somatic pilRNAs in Drosophila: contributions of Loqs-PD and R2D2.

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Milijana Mirkovic-Hösle

Full Text Available Transposable elements are a serious threat for genome integrity and their control via small RNA mediated silencing pathways is an ancient strategy. The fruit fly Drosophila melanogaster has two silencing activities that target transposons: endogenous siRNAs (esiRNAs or endo-siRNAs and Piwi-interacting small RNAs (piRNAs. The biogenesis of endo-siRNAs involves the Dicer-2 co-factors Loqs-PD, which acts predominantly during processing of dsRNA by Dcr-2, and R2D2, which primarily helps to direct siRNAs into the RNA interference effector Ago2. Nonetheless, loss of either protein is not sufficient to produce a phenotype comparable with a dcr-2 mutation. We provide further deep sequencing evidence supporting the notion that R2D2 and Loqs-PD have partially overlapping function. Certain transposons display a preference for either dsRBD-protein during production or loading; this appeared to correlate neither with overall abundance, classification of the transposon or a specific site of genomic origin. The endo-siRNA biogenesis pathway in germline operates according to the same principles as the existing model for the soma, and its impairment does not significantly affect piRNAs. Expanding the analysis, we confirmed the occurrence of somatic piRNA-like RNAs (pilRNAs that show a ping-pong signature. We detected expression of the Piwi-family protein mRNAs only barely above background, indicating that the somatic pilRNAs may arise from a small sub-population of somatic cells that express a functional piRNA pathway.

Remanufacture Systems for Category 1 and 2 Marine Diesel Engines

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EPA maintains a list of remanufacture systems, or “kits”, certified for use with Category 1 and 2 marine diesel engines according to the provisions of 40 CFR Part 1042, Subpart I, and is periodically updated.

Germline transformation of the Mediterranean fruit fly, Ceratitis capitata

International Nuclear Information System (INIS)

McCombs, Susan D.

2000-01-01

Gene transfer methodology for insects was first developed in Drosophila melanogaster Meigen using a transposon-mediated system based on the P element (Spradling and Rubin 1982, Rubin and Spradling 1982). In addition to the P element, three unrelated transposons have been used successfully in genetic transformation of D. melanogaster: hobo (Blackman et al. 1989), Minos (Loukeris et al. 1992), and mariner (Lidholm et al. 1993). Routine gene transfer in Drosophila created a great deal of optimism amongst researchers who sought to employ transgenic techniques in other arthropods. However, what followed were years of consistently disappointing results in other insect species. For example, the P element system was tried unsuccessfully in several species, but was eventually shown to be non-functional outside the genus Drosophila (O'Brochta and Handler 1988). Ensuing research in non-drosophilids emphasised testing of other Drosophila systems and development of transposons isolated from other species. After nearly 15 years of intensive effort, the first successes have only recently been reported. Three Drosophila-derived transposon-based systems: hobo from D. melanogaster, mariner from Drosophila mauritiana Tsacas and David and Minos from Drosophila hydei Sturtevant have produced germline transformation in Drosophila virilis Sturtevant (Gomez and Handler 1997, Lozovskaya et al. 1996), Aedes aegypti L. (Coates et al. 1998), and Ceratitis capitata (Wied.) (Loukeris et al. 1995), respectively. Germline transformation was accomplished with two transposon-based systems from non-drosophilids, Hermes from Musca domestica L. and piggyBac from Trichoplusia ni Huebner in A. aegypti and C. capitata, respectively

Marine Tactical Command and Control System (MTACCS) Field Development System-1 (FDS-1) assessment: Final report

Energy Technology Data Exchange (ETDEWEB)

Avery, L.W.; Hunt, S.T.; Savage, S.F. (Pacific Northwest Lab., Richland, WA (United States)); McLaughlin, P.D.; Shepdard, A.P.; Worl, J.C. (Battelle Seattle Research Center, WA (United States))

1992-04-01

The United State Marine Corps (USMC) is continuing the development and fielding of the Marine Corps Tactical Command and Control System (MTACCS), a system which exists in varying states of development, fielding, or modernization. MTACCS is currently composed of the following components: Tactical Combat Operations System (TCO) for ground command and control (C2), Intelligence Analysis System (IAS) with a Genser terminal connected to a TCO workstation for intelligence C2, Marine Integrated Personnel System (MIPS) and a TCO workstation using the Marine Combat Personnel System (MCPERS) software for personnel C2, Marine Integrated Logistics System (MILOGS) which is composed of the Landing Force Asset Distribution System (LFADS), the Marine Air-Ground Task Force (MAGTF) II, and a TCO terminal using the Marine Combat Logistics System (MCLOG) for logistics C2, Marine Corps Fire Support System (MCFSS) for fire support C2, and Advanced Tactical Air Command Central (ATACC) and the Improved Direct Air Support Central for aviation C2.

Structural and sequence diversity of the transposon Galileo in the Drosophila willistoni genome.

Science.gov (United States)

Gonçalves, Juliana W; Valiati, Victor Hugo; Delprat, Alejandra; Valente, Vera L S; Ruiz, Alfredo

2014-09-13

Galileo is one of three members of the P superfamily of DNA transposons. It was originally discovered in Drosophila buzzatii, in which three segregating chromosomal inversions were shown to have been generated by ectopic recombination between Galileo copies. Subsequently, Galileo was identified in six of 12 sequenced Drosophila genomes, indicating its widespread distribution within this genus. Galileo is strikingly abundant in Drosophila willistoni, a neotropical species that is highly polymorphic for chromosomal inversions, suggesting a role for this transposon in the evolution of its genome. We carried out a detailed characterization of all Galileo copies present in the D. willistoni genome. A total of 191 copies, including 133 with two terminal inverted repeats (TIRs), were classified according to structure in six groups. The TIRs exhibited remarkable variation in their length and structure compared to the most complete copy. Three copies showed extended TIRs due to internal tandem repeats, the insertion of other transposable elements (TEs), or the incorporation of non-TIR sequences into the TIRs. Phylogenetic analyses of the transposase (TPase)-encoding and TIR segments yielded two divergent clades, which we termed Galileo subfamilies V and W. Target-site duplications (TSDs) in D. willistoni Galileo copies were 7- or 8-bp in length, with the consensus sequence GTATTAC. Analysis of the region around the TSDs revealed a target site motif (TSM) with a 15-bp palindrome that may give rise to a stem-loop secondary structure. There is a remarkable abundance and diversity of Galileo copies in the D. willistoni genome, although no functional copies were found. The TIRs in particular have a dynamic structure and extend in different ways, but their ends (required for transposition) are more conserved than the rest of the element. The D. willistoni genome harbors two Galileo subfamilies (V and W) that diverged ~9 million years ago and may have descended from an ancestral

Marine nitrogen cycle

Digital Repository Service at National Institute of Oceanography (India)

Naqvi, S.W.A.

) such as the Marine nitrogen cycle The marine nitrogen cycle. ‘X’ and ‘Y’ are intra-cellular intermediates that do not accumulate in water column. (Source: Codispoti et al., 2001) Page 1 of 3Marine nitrogen cycle - Encyclopedia of Earth 11/20/2006http://www... and nitrous oxide budgets: Moving targets as we enter the anthropocene?, Sci. Mar., 65, 85-105, 2001. Page 2 of 3Marine nitrogen cycle - Encyclopedia of Earth 11/20/2006http://www.eoearth.org/article/Marine_nitrogen_cycle square6 Gruber, N.: The dynamics...

Effects of Biodiesel Blend on Marine Fuel Characteristics for Marine Vessels

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Cherng-Yuan Lin

2013-09-01

Full Text Available Biodiesel produced from vegetable oils, animal fats and algae oil is a renewable, environmentally friendly and clean alternative fuel that reduces pollutants and greenhouse gas emissions in marine applications. This study investigates the influence of biodiesel blend on the characteristics of residual and distillate marine fuels. Adequate correlation equations are applied to calculate the fuel properties of the blended marine fuels with biodiesel. Residual marine fuel RMA has inferior fuel characteristics compared with distillate marine fuel DMA and biodiesel. The flash point of marine fuel RMA could be increased by 20% if blended with 20 vol% biodiesel. The sulfur content of residual marine fuel could meet the requirement of the 2008 MARPOL Annex VI Amendment by blending it with 23.0 vol% biodiesel. In addition, the kinematic viscosity of residual marine fuel could be reduced by 12.9% and the carbon residue by 23.6% if 20 vol% and 25 vol% biodiesel are used, respectively. Residual marine fuel blended with 20 vol% biodiesel decreases its lower heating value by 1.9%. Moreover, the fuel properties of residual marine fuel are found to improve more significantly with biodiesel blending than those of distillate marine fuel.

Italy. Report 1 [Marine Radioecology. Current Research and Future Scope

International Nuclear Information System (INIS)

Argiero, L.

1967-01-01

Present research programme: Studies on radioactive, biological and hydrographic characteristics of Tyrrhenian Sea; Organization of suitable control network of marine water and fauna: systematic measurement of present components of natural and artificial radioactivity of water, sediments and of the marine food chain techniques of sampling; analysis and measurement of particular radioisotopes in marine water. α, β, γ analysis of samples; γ spectrometry, research on 90 Sr and 137 Cs. Determination of concentration factors of radioisotopes which are responsible for contamination of principal components of the flora and fauna in the Tyrrhenian Sea; research of hydrographie factors influencing the distribution of marine radioactivity. Nuclide content of other organisms of the Tyrrhenian flora and fauna

Complete nucleotide sequence of CTX-M-15-plasmids from clinical Escherichia coli isolates: insertional events of transposons and insertion sequences.

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Annemieke Smet

Full Text Available BACKGROUND: CTX-M-producing Escherichia coli strains are regarded as major global pathogens. METHODOLOGY/PRINCIPAL FINDINGS: The nucleotide sequence of three plasmids (pEC_B24: 73801-bp; pEC_L8: 118525-bp and pEC_L46: 144871-bp from Escherichia coli isolates obtained from patients with urinary tract infections and one plasmid (pEC_Bactec: 92970-bp from an Escherichia coli strain isolated from the joint of a horse with arthritis were determined. Plasmid pEC_Bactec belongs to the IncI1 group and carries two resistance genes: bla(TEM-1 and bla(CTX-M-15. It shares more than 90% homology with a previously published bla(CTX-M-plasmid from E. coli of human origin. Plasmid pEC_B24 belongs to the IncFII group whereas plasmids pEC_L8 and pEC_L46 represent a fusion of two replicons of type FII and FIA. On the pEC_B24 backbone, two resistance genes, bla(TEM-1 and bla(CTX-M-15, were found. Six resistance genes, bla(TEM-1, bla(CTX-M-15, bla(OXA-1, aac6'-lb-cr, tetA and catB4, were detected on the pEC_L8 backbone. The same antimicrobial drug resistance genes, with the exception of tetA, were also identified on the pEC_L46 backbone. Genome analysis of all 4 plasmids studied provides evidence of a seemingly frequent transposition event of the bla(CTX-M-15-ISEcp1 element. This element seems to have a preferred insertion site at the tnpA gene of a bla(TEM-carrying Tn3-like transposon, the latter itself being inserted by a transposition event. The IS26-composite transposon, which contains the bla(OXA-1, aac6'-lb-cr and catB4 genes, was inserted into plasmids pEC_L8 and pEC_L46 by homologous recombination rather than a transposition event. Results obtained for pEC_L46 indicated that IS26 also plays an important role in structural rearrangements of the plasmid backbone and seems to facilitate the mobilisation of fragments from other plasmids. CONCLUSIONS: Collectively, these data suggests that IS26 together with ISEcp1 could play a critical role in the evolution of

Marine environment news. Vol. 5, no. 1, September 2007

International Nuclear Information System (INIS)

2007-08-01

Marine Environment Laboratories (MEL) new programmes and strategic outreach efforts (e.g. Mission visits, Newsletter, displays at IAEA's 50th General Conference) have resulted this year in a record number of Marine Technical Co-operation (TC) projects. They increased from 17 in 2006 to 28 in 2007/08 and included 9 regional and 3 interregional projects, now directly benefiting 64 Member States (MS). This is clear evidence of increased interest by Member States to fully utilize the Agency's expertise to better understand and protect their diverse marine environments and resources. I would also like to thank MEL staff for rising to the challenge of supporting these new TC projects. We are also pleased to report that in June 2007, we had a record number of Vienna-based Missions (22 representatives) including the Chairman of the IAEA Board of Governors, the DDG, Mr. W. Burkart, and two Ministers from Monaco visiting MEL in Monaco. The VIP delegates toured MEL's world class laboratories and concluded with a round table discussion on training MS in marine radioactivity and radioecology, on applying new isotopic tools for marine pollution and ocean climate change studies, and on investing in essential equipment at MEL. I am also pleased to report that the IAEA Board of Governors have just approved the replacement of MEL's ageing High Resolution Inductively Coupled Plasma Mass Spectrometer (ICP-MS) now planned for 2008

A protocol for the intercomparison of marine fishery and ecosystem models: Fish-MIP v1.0

Science.gov (United States)

Tittensor, Derek P.; Eddy, Tyler D.; Lotze, Heike K.; Galbraith, Eric D.; Cheung, William; Barange, Manuel; Blanchard, Julia L.; Bopp, Laurent; Bryndum-Buchholz, Andrea; Büchner, Matthias; Bulman, Catherine; Carozza, David A.; Christensen, Villy; Coll, Marta; Dunne, John P.; Fernandes, Jose A.; Fulton, Elizabeth A.; Hobday, Alistair J.; Huber, Veronika; Jennings, Simon; Jones, Miranda; Lehodey, Patrick; Link, Jason S.; Mackinson, Steve; Maury, Olivier; Niiranen, Susa; Oliveros-Ramos, Ricardo; Roy, Tilla; Schewe, Jacob; Shin, Yunne-Jai; Silva, Tiago; Stock, Charles A.; Steenbeek, Jeroen; Underwood, Philip J.; Volkholz, Jan; Watson, James R.; Walker, Nicola D.

2018-04-01

Model intercomparison studies in the climate and Earth sciences communities have been crucial to building credibility and coherence for future projections. They have quantified variability among models, spurred model development, contrasted within- and among-model uncertainty, assessed model fits to historical data, and provided ensemble projections of future change under specified scenarios. Given the speed and magnitude of anthropogenic change in the marine environment and the consequent effects on food security, biodiversity, marine industries, and society, the time is ripe for similar comparisons among models of fisheries and marine ecosystems. Here, we describe the Fisheries and Marine Ecosystem Model Intercomparison Project protocol version 1.0 (Fish-MIP v1.0), part of the Inter-Sectoral Impact Model Intercomparison Project (ISIMIP), which is a cross-sectoral network of climate impact modellers. Given the complexity of the marine ecosystem, this class of models has substantial heterogeneity of purpose, scope, theoretical underpinning, processes considered, parameterizations, resolution (grain size), and spatial extent. This heterogeneity reflects the lack of a unified understanding of the marine ecosystem and implies that the assemblage of all models is more likely to include a greater number of relevant processes than any single model. The current Fish-MIP protocol is designed to allow these heterogeneous models to be forced with common Earth System Model (ESM) Coupled Model Intercomparison Project Phase 5 (CMIP5) outputs under prescribed scenarios for historic (from the 1950s) and future (to 2100) time periods; it will be adapted to CMIP phase 6 (CMIP6) in future iterations. It also describes a standardized set of outputs for each participating Fish-MIP model to produce. This enables the broad characterization of differences between and uncertainties within models and projections when assessing climate and fisheries impacts on marine ecosystems and the

A protocol for the intercomparison of marine fishery and ecosystem models: Fish-MIP v1.0

Directory of Open Access Journals (Sweden)

D. P. Tittensor

2018-04-01

Full Text Available Model intercomparison studies in the climate and Earth sciences communities have been crucial to building credibility and coherence for future projections. They have quantified variability among models, spurred model development, contrasted within- and among-model uncertainty, assessed model fits to historical data, and provided ensemble projections of future change under specified scenarios. Given the speed and magnitude of anthropogenic change in the marine environment and the consequent effects on food security, biodiversity, marine industries, and society, the time is ripe for similar comparisons among models of fisheries and marine ecosystems. Here, we describe the Fisheries and Marine Ecosystem Model Intercomparison Project protocol version 1.0 (Fish-MIP v1.0, part of the Inter-Sectoral Impact Model Intercomparison Project (ISIMIP, which is a cross-sectoral network of climate impact modellers. Given the complexity of the marine ecosystem, this class of models has substantial heterogeneity of purpose, scope, theoretical underpinning, processes considered, parameterizations, resolution (grain size, and spatial extent. This heterogeneity reflects the lack of a unified understanding of the marine ecosystem and implies that the assemblage of all models is more likely to include a greater number of relevant processes than any single model. The current Fish-MIP protocol is designed to allow these heterogeneous models to be forced with common Earth System Model (ESM Coupled Model Intercomparison Project Phase 5 (CMIP5 outputs under prescribed scenarios for historic (from the 1950s and future (to 2100 time periods; it will be adapted to CMIP phase 6 (CMIP6 in future iterations. It also describes a standardized set of outputs for each participating Fish-MIP model to produce. This enables the broad characterization of differences between and uncertainties within models and projections when assessing climate and fisheries impacts on marine ecosystems

Isolation of bacterial extrachromosomal DNA from human dental plaque associated with periodontal disease, using transposon-aided capture (TRACA).

Science.gov (United States)

Warburton, Philip J; Allan, Elaine; Hunter, Stephanie; Ward, John; Booth, Veronica; Wade, William G; Mullany, Peter

2011-11-01

The human oral cavity is host to a complex microbial community estimated to comprise >700 bacterial species, of which at least half are thought to be not yet cultivable in vitro. To investigate the plasmids present in this community, we used a transposon-aided capture system, which allowed the isolation of plasmids from human oral supra- and subgingival plaque samples. Thirty-two novel plasmids and a circular molecule that could be an integrase-generated circular intermediate were isolated. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

A LTR copia retrotransposon and Mutator transposons interrupt Pgip genes in cultivated and wild wheats.

Science.gov (United States)

Di Giovanni, Michela; Cenci, Alberto; Janni, Michela; D'Ovidio, Renato

2008-04-01

Polygalacturonase-inhibiting proteins (PGIPs) are leucine-rich repeat (LRR) proteins involved in plant defence. Wheat pgip genes have been isolated from the B (Tapgip1) and D (Tapgip2) genomes, and now we report the identification of pgip genes from the A genomes of wild and cultivated wheats. By Southern blots and sequence analysis of BAC clones we demonstrated that wheat contains a single copy pgip gene per genome and the one from the A genome, pgip3, is inactivated by the insertion of a long terminal repeat copia retrotranspon within the fourth LRR. We demonstrated also that this retrotransposon insertion is present in Triticum urartu and all the polyploidy wheats assayed, but is absent in T. monococcum (Tmpgip3), suggesting that this insertion took place after the divergence between T. monococcum and T. urartu, but before the formation of the polyploid wheats. We identified also two independent insertion events of new Class II transposable elements, Vacuna, belonging to the Mutator superfamily, that interrupted the Tdipgip1 gene of T. turgidum ssp. dicoccoides. The occurrence of these transposons within the coding region of Tdipgip1 facilitated the mapping of the Pgip locus in the pericentric region of the short arm of chromosome group 7. We speculate that the inactivation of pgip genes are tolerated because of redundancy of PGIP activities in the wheat genome.

In silico characterization of microsatellites in Eucalyptus spp.: abundance, length variation and transposon associations

Directory of Open Access Journals (Sweden)

Edenilson Rabello

2005-01-01

Full Text Available This study assessed the abundance of microsatellites, or simple sequence repeats (SSR, in 19 Eucalyptus EST libraries from FORESTs, containing cDNA sequences from five species: E. grandis, E. globulus, E. saligna, E. urophylla and E. camaldulensis. Overall, a total of 11,534 SSRs and 8,447 SSR-containing sequences (25.5% of total ESTs were identified, with an average of 1 SSR/2.5 kb when considering all motifs and 1 SSR/3.1 kb when mononucleotides were not included. Dimeric repeats were the most abundant (41.03%, followed by trimerics (36.11% and monomerics (19.59%. The most frequent motifs were A/T (87.24% for monomerics, AG/CT (94.44% for dimerics, CCG/CGG (37.87% for trimerics, AAGG/CCTT (18.75% for tetramerics, AGAGG/CCTCT (14.04% for pentamerics and ACGGCG/CGCCGT (6.30% for hexamerics. According to sequence length, Class II or potentially variable markers were the most commonly found, followed by Class III. Two sequences presented high similarity to previously published Eucalyptus sequences from the NCBI database, EMBRA_72 and EMBRA_122. Local blastn search for transposons did not reveal the presence of any transposable elements with a cut-off value of 10-50. The large number of microsatellites identified will contribute to the refinement of marker-assisted mapping and to the discovery of novel markers for virtually all genes of economic interest.

Time-Resolved Transposon Insertion Sequencing Reveals Genome-Wide Fitness Dynamics during Infection.

Science.gov (United States)

Yang, Guanhua; Billings, Gabriel; Hubbard, Troy P; Park, Joseph S; Yin Leung, Ka; Liu, Qin; Davis, Brigid M; Zhang, Yuanxing; Wang, Qiyao; Waldor, Matthew K

2017-10-03

Transposon insertion sequencing (TIS) is a powerful high-throughput genetic technique that is transforming functional genomics in prokaryotes, because it enables genome-wide mapping of the determinants of fitness. However, current approaches for analyzing TIS data assume that selective pressures are constant over time and thus do not yield information regarding changes in the genetic requirements for growth in dynamic environments (e.g., during infection). Here, we describe structured analysis of TIS data collected as a time series, termed pattern analysis of conditional essentiality (PACE). From a temporal series of TIS data, PACE derives a quantitative assessment of each mutant's fitness over the course of an experiment and identifies mutants with related fitness profiles. In so doing, PACE circumvents major limitations of existing methodologies, specifically the need for artificial effect size thresholds and enumeration of bacterial population expansion. We used PACE to analyze TIS samples of Edwardsiella piscicida (a fish pathogen) collected over a 2-week infection period from a natural host (the flatfish turbot). PACE uncovered more genes that affect E. piscicida 's fitness in vivo than were detected using a cutoff at a terminal sampling point, and it identified subpopulations of mutants with distinct fitness profiles, one of which informed the design of new live vaccine candidates. Overall, PACE enables efficient mining of time series TIS data and enhances the power and sensitivity of TIS-based analyses. IMPORTANCE Transposon insertion sequencing (TIS) enables genome-wide mapping of the genetic determinants of fitness, typically based on observations at a single sampling point. Here, we move beyond analysis of endpoint TIS data to create a framework for analysis of time series TIS data, termed pattern analysis of conditional essentiality (PACE). We applied PACE to identify genes that contribute to colonization of a natural host by the fish pathogen

Influence of marine current on vertical migration of Pb in marine bay

Science.gov (United States)

Yu, Chen; Hong, Ai; Danfeng, Yang; Huijuan, Zhao; Dongfang, Yang

2018-02-01

This paper analyzed that vertical migration of Pb contents waters in Jiaozhou Bay, and revealed the influence of marine current on vertical migration process. Results showed that Pb contents in bottom waters of Jiaozhou Bay in April and July 1988 were 1.49-18.53 μg L-1 and 12.68/-27.64 μg L-1, respectively. The pollution level of Pb in bottom waters was moderate to heavy, and were showing temporal variations and spatial heterogeneity. The vertical migration process of Pb in April 1988 included a drifting process from the southwest to the north by means of the marine current was rapid in this region. The vertical migration process of Pb in July 1988 in the open waters included no drifting process since the flow rate of marine current was relative low in this region. The vertical migration process of Pb was jointly determined by vertical water’s effect, source input and water exchange, and the influence of marine current on the vertical migration of Pb in marine bay was significant.

Marine environment news. Vol. 3, no. 1, May 2005

International Nuclear Information System (INIS)

2005-06-01

This issue presents a guest article on selective biomagnification of metals and radionuclides in marine food chains. Major news items include that MEL joins the international study in South Pacific, inter-agency collaboration in the study of marine pollution, IAEA-UNESCO coordinate studies on submarine ground water discharge and collaborative research on nuclear and isotopic studies of the El Nino phenomenon launched in 2004

Marine environment news. Vol. 3, no. 1, May 2005

Energy Technology Data Exchange (ETDEWEB)

NONE

2005-06-01

This issue presents a guest article on selective biomagnification of metals and radionuclides in marine food chains. Major news items include that MEL joins the international study in South Pacific, inter-agency collaboration in the study of marine pollution, IAEA-UNESCO coordinate studies on submarine ground water discharge and collaborative research on nuclear and isotopic studies of the El Nino phenomenon launched in 2004.

Scavenging of radionuclides in the marine environment, (1)

International Nuclear Information System (INIS)

Mahara, Yasunori

1989-01-01

It is very important to predict diffusion, scavenging and accumulation of the long lives radionuclides which may be discharged from the reprocessing plant in the marine environment, for the purpose of polishing up methods of the radiation does estimation to the high quality stage. This study reports that distribution and behavior of transuranic elements, which are extremely harmful for the human beings and are discharged probably from the reprocessing plant, are investigated in both the survey of bibliography and the in-situ observation. Results of the field observation on the distribution of transuranic elements in the marine show that plutonium and americium are easily scavenged from the sea water and are accumulated on the sea bottom. Transuranic elements, which are originated from fallout and are discharged from the reprocessing plant, have generally the similar distribution in the marine and the same chemical behavior. This facts suggest that the fallout data which are probably and easily collected in the world are available for fabrication of the scavenging model for transuranic elements discharged from the reprocessing plants. (author)

2007 Joint Service Power Expo: Power and Energy Independence for Warfighters

Science.gov (United States)

2007-04-26

longitudinal, and lateral directions: - 55-ton overhead chain hoist - Remotely controlled 60-ton hydraulic load application system - Airbag inflation system...Step Apply & Load Dump Inductive Load Start Short Circuit Thermal Limits Step Load Steady State Thermal Loading 0 20 40 60 80 100 120 140 0 1.000...15 ton payload primary and secondary roads • MTVR Based Variants – Cargo, Dump Truck, Wrecker, HIMARS Re-Supply Vehicle, Tractor, LHS (load

Development of Transgenic Cloned Pig Models of Skin Inflammation by DNA Transposon-Directed Ectopic Expression of Human β1 and α2 Integrin

Science.gov (United States)

Staunstrup, Nicklas Heine; Madsen, Johannes; Primo, Maria Nascimento; Li, Juan; Liu, Ying; Kragh, Peter M.; Li, Rong; Schmidt, Mette; Purup, Stig; Dagnæs-Hansen, Frederik; Svensson, Lars; Petersen, Thomas K.; Callesen, Henrik; Bolund, Lars; Mikkelsen, Jacob Giehm

2012-01-01

Integrins constitute a superfamily of transmembrane signaling receptors that play pivotal roles in cutaneous homeostasis by modulating cell growth and differentiation as well as inflammatory responses in the skin. Subrabasal expression of integrins α2 and/or β1 entails hyperproliferation and aberrant differentiation of keratinocytes and leads to dermal and epidermal influx of activated T-cells. The anatomical and physiological similarities between porcine and human skin make the pig a suitable model for human skin diseases. In efforts to generate a porcine model of cutaneous inflammation, we employed the Sleeping Beauty DNA transposon system for production of transgenic cloned Göttingen minipigs expressing human β1 or α2 integrin under the control of a promoter specific for subrabasal keratinocytes. Using pools of transgenic donor fibroblasts, cloning by somatic cell nuclear transfer was utilized to produce reconstructed embryos that were subsequently transferred to surrogate sows. The resulting pigs were all transgenic and harbored from one to six transgene integrants. Molecular analyses on skin biopsies and cultured keratinocytes showed ectopic expression of the human integrins and localization within the keratinocyte plasma membrane. Markers of perturbed skin homeostasis, including activation of the MAPK pathway, increased expression of the pro-inflammatory cytokine IL-1α, and enhanced expression of the transcription factor c-Fos, were identified in keratinocytes from β1 and α2 integrin-transgenic minipigs, suggesting the induction of a chronic inflammatory phenotype in the skin. Notably, cellular dysregulation obtained by overexpression of either β1 or α2 integrin occurred through different cellular signaling pathways. Our findings mark the creation of the first cloned pig models with molecular markers of skin inflammation. Despite the absence of an overt psoriatic phenotype, these animals may possess increased susceptibility to severe skin damage

Transposable elements in phytopathogenic Verticillium spp.: insights into genome evolution and inter- and intra-specific diversification

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Amyotte Stefan G

2012-07-01

Full Text Available Abstract Background Verticillium dahliae (Vd and Verticillium albo-atrum (Va are cosmopolitan soil fungi causing very disruptive vascular diseases on a wide range of crop plants. To date, no sexual stage has been identified in either microorganism suggesting that somatic mutation is a major force in generating genetic diversity. Whole genome comparative analysis of the recently sequenced strains VdLs.17 and VaMs.102 revealed that non-random insertions of transposable elements (TEs have contributed to the generation of four lineage-specific (LS regions in VdLs.17. Results We present here a detailed analysis of Class I retrotransposons and Class II “cut-and-paste” DNA elements detected in the sequenced Verticillium genomes. We report also of their distribution in other Vd and Va isolates from various geographic origins. In VdLs.17, we identified and characterized 56 complete retrotransposons of the Gypsy-, Copia- and LINE-like types, as well as 34 full-length elements of the “cut-and-paste” superfamilies Tc1/mariner, Activator and Mutator. While Copia and Tc1/mariner were present in multiple identical copies, Activator and Mutator sequences were highly divergent. Most elements comprised complete ORFs, had matching ESTs and showed active transcription in response to stress treatment. Noticeably, we found evidences of repeat-induced point mutation (RIP only in some of the Gypsy retroelements. While Copia-, Gypsy- and Tc1/mariner-like transposons were prominent, a large variation in presence of the other types of mobile elements was detected in the other Verticillium spp. strains surveyed. In particular, neither complete nor defective “cut-and-paste” TEs were found in VaMs.102. Conclusions Copia-, Gypsy- and Tc1/mariner-like transposons are the most wide-spread TEs in the phytopathogens V. dahliae and V. albo-atrum. In VdLs.17, we identified several retroelements and “cut-and-paste” transposons still potentially active. Some of these

Genotype-dependent Burst of Transposable Element Expression in Crowns of Hexaploid Wheat (Triticum aestivum L. during Cold Acclimation

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Debbie Laudencia-Chingcuanco

2012-01-01

Full Text Available The expression of 1,613 transposable elements (TEs represented in the Affymetrix Wheat Genome Chip was examined during cold treatment in crowns of four hexaploid wheat genotypes that vary in tolerance to cold and in flowering time. The TE expression profiles showed a constant level of expression throughout the experiment in three of the genotypes. In winter Norstar, the most cold-hardy of the four genotypes, a subset of the TEs showed a burst of expression after vernalization saturation was achieved. About 47% of the TEs were expressed, and both Class I (retrotransposons and Class II (DNA transposons types were well represented. Gypsy and Copia were the most represented among the retrotransposons while CACTA and Mariner were the most represented DNA transposons. The data suggests that the Vrn-A1 region plays a role in the stage-specific induction of TE expression in this genotype.

Diversity of the tetracycline resistance gene tet(M) and identification of Tn916- and Tn5801-like (Tn6014) transposons in Staphylococcus aureus from humans and animals

DEFF Research Database (Denmark)

de Vries, Lisbeth Elvira; Christensen, H.; Skov, R. L.

2009-01-01

To analyse the sequence diversity of the tetracycline resistance gene tet(M) in Staphylococcus aureus from humans and animals and to determine mobile elements associated with tet(M) in S. aureus. In total, 205 tetracycline-resistant isolates were screened for tet(M) by PCR. tet(M) genes were...... sequenced and compared with tet(M) deposited in GenBank. Based on phylogenetic analysis isolates were screened for Tn916- and Tn5801-like xis/int genes, and transposons were confirmed by linking PCR. spa typing was performed and selected isolates were used as donors in a filter mating experiment. Forty......-one isolates (21.3%, 60.7%, 2.6% and 4.4% of the human, pig, poultry and cattle isolates, respectively) were tet(M) positive. tet(M) was located on Tn5801-like and Tn916-like transposons in humans and on a specific Tn916-like element in animals. Human isolates were of different spa types (t034, t008, t037, t...

Radiation and transposon-induced genetic damage in Drosophila melanogaster

International Nuclear Information System (INIS)

Balter, H.; Griffith, C.S.; American Museum of Natural History, New York; Margulies, L.

1992-01-01

The interaction of X-ray-induced and transposon-induced damage was investigated in P-M hybrid dysgenesis in Drosophila melanogaster. X-ray dose-response of 330-1320 rad was monitored for sterility, fecundicity and partial X/Y chromosome loss among F 2 progeny derived from dysgenic cross of M strain females xP strain males (cross A) and its reciprocal (cross B), using a weaker and the standard Harwich P strain subline. The synergistic effect of P element activity and X-rays on sterility was observed only in cross A hybrids and the dose-response was nonlinear in hybrids derived from the strong standard reference Harwich subline, H W . This finding suggests that lesions induced by both mutator systems which produce the synergistic effects are 2-break events. Effect of increasing dose on the decline of fecundicity was synergistic, but linear, in hybrids of either subline. There was no interaction evident and thus no synergism in X/Y nondisjunction and partial Y chromosome loss measured by the loss of the B s marker alone or together with the y + marker. Interaction was detected in the loss of the y + marker alone from the X and Y chromosomes. The possible three-way interaction of X-rays (660 rad), post-replication repair deficiency and P elements mobility was assessed by measuring transmission distortion in dysgenic males derived from the Π 2 P strain. (author). 38 refs.; 5 tabs

Yugoslavia. Report 1 [Marine Radioecology. Current Research and Future Scope

Energy Technology Data Exchange (ETDEWEB)

Kečkeš, S.; Pučar, Z. [Laboratory of Marine Radiobiology and Laboratory for Electromigration, Institute ' ' Ruder Boskovic' ' , Rovinj and Zagreb, Yugoslavia (Croatia)

1967-03-15

Present research programme (long-term): Transport of various radiqnuclides in marine environment. Uptake, loss and accumulation of radionuclides in selected marine biota. Study of the physico-chemical forms of various radionuclides in sea water. Tracer experiments on the uptake and loss rate in biota. Electromigration techniques for the characterization of the physico-chemical forms of radionuclides.

Horizontal transfer of transposons between and within crustaceans and insects.

Science.gov (United States)

Dupeyron, Mathilde; Leclercq, Sébastien; Cerveau, Nicolas; Bouchon, Didier; Gilbert, Clément

2014-01-29

Horizontal transfer of transposable elements (HTT) is increasingly appreciated as an important source of genome and species evolution in eukaryotes. However, our understanding of HTT dynamics is still poor in eukaryotes because the diversity of species for which whole genome sequences are available is biased and does not reflect the global eukaryote diversity. In this study we characterized two Mariner transposable elements (TEs) in the genome of several terrestrial crustacean isopods, a group of animals particularly underrepresented in genome databases. The two elements have a patchy distribution in the arthropod tree and they are highly similar (>93% over the entire length of the element) to insect TEs (Diptera and Hymenoptera), some of which were previously described in Ceratitis rosa (Crmar2) and Drosophila biarmipes (Mariner-5_Dbi). In addition, phylogenetic analyses and comparisons of TE versus orthologous gene distances at various phylogenetic levels revealed that the taxonomic distribution of the two elements is incompatible with vertical inheritance. We conclude that the two Mariner TEs each underwent at least three HTT events. Both elements were transferred once between isopod crustaceans and insects and at least once between isopod crustacean species. Crmar2 was also transferred between tephritid and drosophilid flies and Mariner-5 underwent HT between hymenopterans and dipterans. We demonstrate that these various HTTs took place recently (most likely within the last 3 million years), and propose iridoviruses and/or Wolbachia endosymbionts as potential vectors of these transfers.

Genome-wide transposon mutagenesis of Proteus mirabilis: Essential genes, fitness factors for catheter-associated urinary tract infection, and the impact of polymicrobial infection on fitness requirements

Science.gov (United States)

Smith, Sara N.; Zhao, Lili; Wu, Weisheng

2017-01-01

The Gram-negative bacterium Proteus mirabilis is a leading cause of catheter-associated urinary tract infections (CAUTIs), which are often polymicrobial. Numerous prior studies have uncovered virulence factors for P. mirabilis pathogenicity in a murine model of ascending UTI, but little is known concerning pathogenesis during CAUTI or polymicrobial infection. In this study, we utilized five pools of 10,000 transposon mutants each and transposon insertion-site sequencing (Tn-Seq) to identify the full arsenal of P. mirabilis HI4320 fitness factors for single-species versus polymicrobial CAUTI with Providencia stuartii BE2467. 436 genes in the input pools lacked transposon insertions and were therefore concluded to be essential for P. mirabilis growth in rich medium. 629 genes were identified as P. mirabilis fitness factors during single-species CAUTI. Tn-Seq from coinfection with P. stuartii revealed 217/629 (35%) of the same genes as identified by single-species Tn-Seq, and 1353 additional factors that specifically contribute to colonization during coinfection. Mutants were constructed in eight genes of interest to validate the initial screen: 7/8 (88%) mutants exhibited the expected phenotypes for single-species CAUTI, and 3/3 (100%) validated the expected phenotypes for polymicrobial CAUTI. This approach provided validation of numerous previously described P. mirabilis fitness determinants from an ascending model of UTI, the discovery of novel fitness determinants specifically for CAUTI, and a stringent assessment of how polymicrobial infection influences fitness requirements. For instance, we describe a requirement for branched-chain amino acid biosynthesis by P. mirabilis during coinfection due to high-affinity import of leucine by P. stuartii. Further investigation of genes and pathways that provide a competitive advantage during both single-species and polymicrobial CAUTI will likely provide robust targets for therapeutic intervention to reduce P. mirabilis

Genome-wide transposon mutagenesis of Proteus mirabilis: Essential genes, fitness factors for catheter-associated urinary tract infection, and the impact of polymicrobial infection on fitness requirements.

Science.gov (United States)

Armbruster, Chelsie E; Forsyth-DeOrnellas, Valerie; Johnson, Alexandra O; Smith, Sara N; Zhao, Lili; Wu, Weisheng; Mobley, Harry L T

2017-06-01

The Gram-negative bacterium Proteus mirabilis is a leading cause of catheter-associated urinary tract infections (CAUTIs), which are often polymicrobial. Numerous prior studies have uncovered virulence factors for P. mirabilis pathogenicity in a murine model of ascending UTI, but little is known concerning pathogenesis during CAUTI or polymicrobial infection. In this study, we utilized five pools of 10,000 transposon mutants each and transposon insertion-site sequencing (Tn-Seq) to identify the full arsenal of P. mirabilis HI4320 fitness factors for single-species versus polymicrobial CAUTI with Providencia stuartii BE2467. 436 genes in the input pools lacked transposon insertions and were therefore concluded to be essential for P. mirabilis growth in rich medium. 629 genes were identified as P. mirabilis fitness factors during single-species CAUTI. Tn-Seq from coinfection with P. stuartii revealed 217/629 (35%) of the same genes as identified by single-species Tn-Seq, and 1353 additional factors that specifically contribute to colonization during coinfection. Mutants were constructed in eight genes of interest to validate the initial screen: 7/8 (88%) mutants exhibited the expected phenotypes for single-species CAUTI, and 3/3 (100%) validated the expected phenotypes for polymicrobial CAUTI. This approach provided validation of numerous previously described P. mirabilis fitness determinants from an ascending model of UTI, the discovery of novel fitness determinants specifically for CAUTI, and a stringent assessment of how polymicrobial infection influences fitness requirements. For instance, we describe a requirement for branched-chain amino acid biosynthesis by P. mirabilis during coinfection due to high-affinity import of leucine by P. stuartii. Further investigation of genes and pathways that provide a competitive advantage during both single-species and polymicrobial CAUTI will likely provide robust targets for therapeutic intervention to reduce P. mirabilis

Identification of genetic loci required for Campylobacter resistance to fowlicidin-1, a chicken host defense peptide

Directory of Open Access Journals (Sweden)

Ky Van Hoang

2012-03-01

Full Text Available Antimicrobial peptides (AMPs are critical components of host defense limiting bacterial infections at the gastrointestinal mucosal surface. Bacterial pathogens have co-evolved with host innate immunity and developed means to counteract the effect of endogenous AMPs. However, molecular mechanisms of AMP resistance in Campylobacter, an important human food borne pathogen with poultry as a major reservoir, are still largely unknown. In this study, random transposon mutagenesis and targeted site-directed mutagenesis approaches were used to identify genetic loci contributing Campylobacter resistance to fowlicidin-1, a chicken AMP belonging to cathelicidin family. An efficient transposon mutagenesis approach (EZ::TNTM Transposome in conjunction with a microtiter plate screening identified three mutants whose susceptibilities to fowlicidin-1 were significantly increased. Backcrossing of the transposon mutations into parent strain confirmed that the AMP-sensitive phenotype in each mutant was linked to the specific transposon insertion. Direct sequencing showed that these mutants have transposon inserted in the genes encoding two-component regulator CbrR, transporter CjaB, and putative trigger factor Tig. Genomic analysis also revealed an operon (Cj1580c-1584c that is homologous to sapABCDF, an operon conferring resistance to AMP in other pathogens. Insertional inactivation of Cj1583c (sapB significantly increased susceptibility of Campylobacter to fowlicidin-1. The sapB as well as tig and cjaB mutants were significantly impaired in their ability to compete with their wild-type strain 81-176 to colonize the chicken cecum. Together, this study identified four genetic loci in Campylobacter that will be useful for characterizing molecular basis of Campylobacter resistance to AMPs, a significant knowledge gap in Campylobacter pathogenesis.

Transposon mutagenesis identifies novel genes associated with Staphylococcus aureus persister formation

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Wang ewenjie

2015-12-01

Full Text Available Pathogenic bacterial persisters are responsible for the recalcitrance of chronic and persistent infections to antimicrobial therapy. Although the mechanisms of persister formation and survival have been widely studied in Escherichia coli, persistence mechanisms in S. aureus remain largely unknown. Here, we screened a transposon mutant library of a clinical methicillin-resistant Staphylococcus aureus（MRSA）strain, USA500 (ST8, under antibiotic pressure and identified 13 genes whose insertion mutations resulted in a defect in persistence. These candidate genes were further confirmed by evaluating the survival of the mutants upon exposure to levofloxacin and several other stress conditions. We found 13 insertion mutants with significantly lower persister numbers under several stress conditions, including sdhA, sdhB, ureG, mnhG1, fbaA, ctaB, clpX, parE, HOU_0223, HOU_0587, HOU_2091, HOU_2315 and HOU_2346, which mapped into pathways of oxidative phosphorylation, TCA cycle, glycolysis, cell cycle and ABC transporters, suggesting that these genes and pathways may play an important role in persister formation and survival. The newly constructed knockout strains of ureG, sdhA and sdhB and their complemented strains were also tested for defect in persisters following exposure to levofloxacin and several other stress conditions. The results from these experiments were consistent with the screening results, which indicated that deletion of these genes in MRSA USA500 leads to persister defect. These findings provide novel insights into the mechanisms of persister formation and survival in S. aureus and offer new targets for the development of persister-directed antibiotics for the improved treatment of chronic and persistent infections.

Power Projection: The Current and Future Relevance of the M1A1Tank to Marine Corps Capabilities

Science.gov (United States)

2002-01-01

Kenneth W. Estes, Marines under Armor : The Marine Corps and the Armored Fighting Vehicle, 1916-2000 (Annapolis, MD: Naval institute Press, 2000), 183...is the M2 heavy barrel machine gun that has been in the U.S. inventory for decades. It can be fired from under armor and has a range of 1800 meters...Accessed 9 January 2002. Estes, Kenneth W. Marines under Armor : The Marine Corps and the Armored Fighting Vehicle, 1916-2000. Annapolis, MD

50 CFR 14.18 - Marine mammals.

Science.gov (United States)

2010-10-01

... 50 Wildlife and Fisheries 1 2010-10-01 2010-10-01 false Marine mammals. 14.18 Section 14.18....18 Marine mammals. Any person subject to the jurisdiction of the United States who has lawfully taken a marine mammal on the high seas and who is authorized to import such marine mammal in accordance...

Role of transposon-derived small RNAs in the interplay between genomes and parasitic DNA in rice.

Directory of Open Access Journals (Sweden)

Misuzu Nosaka

2012-09-01

Full Text Available RNA silencing is a defense system against "genomic parasites" such as transposable elements (TE, which are potentially harmful to host genomes. In plants, transcripts from TEs induce production of double-stranded RNAs (dsRNAs and are processed into small RNAs (small interfering RNAs, siRNAs that suppress TEs by RNA-directed DNA methylation. Thus, the majority of TEs are epigenetically silenced. On the other hand, most of the eukaryotic genome is composed of TEs and their remnants, suggesting that TEs have evolved countermeasures against host-mediated silencing. Under some circumstances, TEs can become active and increase in copy number. Knowledge is accumulating on the mechanisms of TE silencing by the host; however, the mechanisms by which TEs counteract silencing are poorly understood. Here, we show that a class of TEs in rice produces a microRNA (miRNA to suppress host silencing. Members of the microRNA820 (miR820 gene family are located within CACTA DNA transposons in rice and target a de novo DNA methyltransferase gene, OsDRM2, one of the components of epigenetic silencing. We confirmed that miR820 negatively regulates the expression of OsDRM2. In addition, we found that expression levels of various TEs are increased quite sensitively in response to decreased OsDRM2 expression and DNA methylation at TE loci. Furthermore, we found that the nucleotide sequence of miR820 and its recognition site within the target gene in some Oryza species have co-evolved to maintain their base-pairing ability. The co-evolution of these sequences provides evidence for the functionality of this regulation. Our results demonstrate how parasitic elements in the genome escape the host's defense machinery. Furthermore, our analysis of the regulation of OsDRM2 by miR820 sheds light on the action of transposon-derived small RNAs, not only as a defense mechanism for host genomes but also as a regulator of interactions between hosts and their parasitic elements.

26 CFR 1.821-3 - Tax on mutual insurance companies other than life or marine or fire insurance companies subject...

Science.gov (United States)

2010-04-01

... life or marine or fire insurance companies subject to the tax imposed by section 831. 1.821-3 Section 1... and Other Than Fire Or Flood Insurance Companies Which Operate on Basis of Perpetual Policies Or Premium Deposits) § 1.821-3 Tax on mutual insurance companies other than life or marine or fire insurance...

Mutations in the Histone-Like Nucleoid Structuring Regulatory Gene (hns) Decrease the Adherence of Shiga Toxin-Producing Escherichia coli 091:H21 Strain B2F1 to Human Colonic Epithelial Cells and Increase the Production of Hemolysin

National Research Council Canada - National Science Library

Scott, Maria

1999-01-01

...:H21 to human colonic epithelial cells. Transposon mutagenesis of B2F1 was accomplished with the mini-Tn5phoACm mobile element and a mutant bank of B2F1 colonies that carried putative in-frame PhoA-positive transposon insertions was isolated...

Origin of marine planktonic cyanobacteria.

Science.gov (United States)

Sánchez-Baracaldo, Patricia

2015-12-01

Marine planktonic cyanobacteria contributed to the widespread oxygenation of the oceans towards the end of the Pre-Cambrian and their evolutionary origin represents a key transition in the geochemical evolution of the Earth surface. Little is known, however, about the evolutionary events that led to the appearance of marine planktonic cyanobacteria. I present here phylogenomic (135 proteins and two ribosomal RNAs), Bayesian relaxed molecular clock (18 proteins, SSU and LSU) and Bayesian stochastic character mapping analyses from 131 cyanobacteria genomes with the aim to unravel key evolutionary steps involved in the origin of marine planktonic cyanobacteria. While filamentous cell types evolved early on at around 2,600-2,300 Mya and likely dominated microbial mats in benthic environments for most of the Proterozoic (2,500-542 Mya), marine planktonic cyanobacteria evolved towards the end of the Proterozoic and early Phanerozoic. Crown groups of modern terrestrial and/or benthic coastal cyanobacteria appeared during the late Paleoproterozoic to early Mesoproterozoic. Decrease in cell diameter and loss of filamentous forms contributed to the evolution of unicellular planktonic lineages during the middle of the Mesoproterozoic (1,600-1,000 Mya) in freshwater environments. This study shows that marine planktonic cyanobacteria evolved from benthic marine and some diverged from freshwater ancestors during the Neoproterozoic (1,000-542 Mya).

A combination of quantitative marinating and Maillard reaction to enhance volatile flavor in Chinese marinated chicken.

Science.gov (United States)

Wei, Xiuli; Wang, Chunqing; Zhang, Chunhui; Li, Xia; Wang, Jinzhi; Li, Hai; Tang, Chunhong

2017-02-01

A combination of quantitative marinating and Maillard reaction was investigated by adding d-xylose, l-cysteine and thiamine to the marinated brine of quantitative marinating, which was expected to enhance the volatile flavor of Chinese marinated chicken. Response surface methodology was used to optimize parameters, in which response was sensory evaluation scores of marinated chicken. A Box-Behnken center design was applied to the optimized added contents. The optimized contents were d-xylose (1-5‰), l-cysteine (1-5‰) and thiamine (1-3‰). Analysis of variance indicated that a second-order polynomial equation could predict the experimental data well (R 2  = 0.94), and sensory evaluation scores were significantly affected by the added amount of d-xylose, l-cysteine and thiamine. The optimal conditions that maximized the sensory evaluation score of Chinese marinated chicken were found to be 4.96‰ d-xylose, 2.28‰ l-cysteine and 2.66‰ thiamine (w/w). Given these optimal conditions, a number of meat-like flavor compounds such as 2-pentyl-furan, benzothiazole and 4-methyl-5-thiazoleethanol were identified by gas chromatographic-mass spectrometric analysis. Our results suggested that a combination of quantitative marinating and Maillard reaction might be a promising method to enhance the volatile flavor, especially meat-like flavor, of Chinese marinated chicken. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

78 FR 52135 - Takes of Marine Mammals Incidental to Specified Activities; Taking Marine Mammals Incidental To...

Science.gov (United States)

2013-08-22

...--Marine Mammal Density Estimates Density Species (animals/km \\2\\) Bottlenose dolphin \\1\\ 0.455 Atlantic... criteria and thresholds in a final rule on the unintentional taking of marine animals occurring incidental... analysis assumed the marine species populations were 100 percent small animals. The criterion with the...

Transposon mutagenesis reveals differential pathogenesis of Ralstonia solanacearum on tomato and Arabidopsis.

Science.gov (United States)

Lin, Yu-Mei; Chou, I-Chun; Wang, Jaw-Fen; Ho, Fang-I; Chu, Yu-Ju; Huang, Pei-Cheng; Lu, Der-Kang; Shen, Hwei-Ling; Elbaz, Mounira; Huang, Shu-Mei; Cheng, Chiu-Ping

2008-09-01

Ralstonia solanacearum causes a deadly wilting disease on a wide range of crops. To elucidate pathogenesis of this bacterium in different host plants, we set out to identify R. solanacearum genes involved in pathogenesis by screening random transposon insertion mutants of a highly virulent strain, Pss190, on tomato and Arabidopsis thaliana. Mutants exhibiting various decreased virulence levels on these two hosts were identified. Sequence analysis showed that most, but not all, of the identified pathogenesis genes are conserved among distinct R. solanacearum strains. A few of the disrupted loci were not reported previously as being involved in R. solanacearum pathogenesis. Notably, a group of mutants exhibited differential pathogenesis on tomato and Arabidopsis. These results were confirmed by characterizing allelic mutants in one other R. solanacearum strain of the same phylotype. The significantly decreased mutants' colonization in Arabidopsis was found to be correlated with differential pathogenesis on these two plants. Differential requirement of virulence genes suggests adaptation of this bacterium in different host environments. Together, this study reveals commonalities and differences of R. solanacearum pathogenesis on single solanaceous and nonsolanaceous hosts, and provides important new insights into interactions between R. solanacearum and different host plants.

Xanthomonas oryzae pv oryzae the Causal Agent of Bacterial Leaf Blight of rice: Isolation, Characterization, and Study of Transposon Mutagenesis

Directory of Open Access Journals (Sweden)

Abdjad Asih Nawangsih

2011-04-01

Full Text Available Xanthomonas oryzae pv oryzae the Causal Agent of Bacterial Leaf Blight of rice: Isolation, Characterization, and Study of Transposon Mutagenesis. X. oryzae pv. oryzae (Xoo causes bacterial leaf blight (BLB of rice (Oryza sativa L., a major disease that constrains production of the staple crop in many countries of the world. Identification of X. oryzae pv. oryzae (Xoo was conducted based on the disease symptoms, pathogenicity, morphological, physiological, and genetic characteristics of bacterial cultures isolated from the infected plants. Fifty bacterial isolates predicted as Xoo have been successfully isolated. They are aerobic, rod shaped, and Gram negative bacteria. The isolates were evaluated for their hypersensitivity in tobacco and pathogenicity in rice plant. Fifty isolates induced hypersensitive reaction in tobacco and showed pathogenicity symptom in rice in different length. Based on physiological test, hypersensitivity and pathogenicity reactions, three bacterial isolates strongly predicted as Xoo, i.e. STG21, STG42, and STG46, were non indole formation, non pigment fluorescent, hydrolyzed casein, catalase activity positive, but negative oxidase. Partial sequencing of 16S rRNA genes of STG21 and STG42 showed 80% and 82% homology with X. oryzae, respectively, while STG46 showed 84% homology with X. campestris. Mini-Tn5 transposon mutagenesis of STG21 generated one of the mutants (M5 lossed it’s ability to induce hypersensitive reaction in tobacco plant and deficient in pathogenicity on rice. The lesion length of rice leaf caused by the mutant M5 decreased up to 80%.

Genome-Wide Analysis of Salicylate and Dibenzofuran Metabolism in Sphingomonas wittichii RW1

Directory of Open Access Journals (Sweden)

Edith eCoronado

2012-08-01

Full Text Available Sphingomonas wittichii RW1 is a bacterium isolated for its ability to degrade the xenobiotic compounds dibenzodioxin and dibenzofuran (DBF. A number of genes involved in DBF degradation have been previously characterized, such as the dxn cluster, dbfB, and the electron transfer components fdx1, fdx3 and redA2. Here we use a combination of whole genome transcriptome analysis and transposon library screening to characterize RW1 catabolic and other genes implicated in the reaction to or degradation of DBF. To detect differentially expressed genes upon exposure to DBF, we applied three different growth exposure experiments, using either short DBF exposures to actively growing cells or growing them with DBF as sole carbon and energy source. Genome-wide gene expression was examined using a custom-made microarray. In addition, proportional abundance determination of transposon insertions in RW1 libraries grown on salicylate or DBF by ultra-high throughput sequencing was used to infer genes whose interruption caused a fitness loss for growth on DBF. Expression patterns showed that batch and chemostat growth conditions, and short or long exposure of cells to DBF produced very different responses. Numerous other uncharacterized catabolic gene clusters putatively involved in aromatic compound metabolism increased expression in response to DBF. In addition, only very few transposon insertions completely abolished growth on DBF. Some of those (e.g., in dxnA1 were expected, whereas others (in a gene cluster for phenylacetate degradation were not. Both transcriptomic data and transposon screening suggest operation of multiple redundant and parallel aromatic pathways, depending on DBF exposure. In addition, increased expression of other non-catabolic genes suggests that during initial exposure, S. wittichii RW1 perceives DBF as a stressor, whereas after longer exposure, the compound is recognized as a carbon source and metabolized using several pathways in

ZBED6, a novel transcription factor derived from a domesticated DNA transposon regulates IGF2 expression and muscle growth

DEFF Research Database (Denmark)

Markljung, Ellen; Jiang, Lin; Jaffe, Jacob D

2009-01-01

and find that the protein, named ZBED6, is previously unknown, specific for placental mammals, and derived from an exapted DNA transposon. Silencing of Zbed6 in mouse C2C12 myoblasts affected Igf2 expression, cell proliferation, wound healing, and myotube formation. Chromatin immunoprecipitation (Ch......, including development and transcriptional regulation. The phenotypic effects in mutant pigs and ZBED6-silenced C2C12 myoblasts, the extreme sequence conservation, its nucleolar localization, the broad tissue distribution, and the many target genes with essential biological functions suggest that ZBED6...... is an important transcription factor in placental mammals, affecting development, cell proliferation, and growth....

The Use of Stimulable Bioluminescence from Marine Dinoflagellates as a Means of Detecting Toxicity in the Marine Environment

Science.gov (United States)

1993-04-01

FROM MARINE PR: ME65 DINOFLAGELLATES AS A MEANS OF DETECTING TOXICITY IN THE PE: 060372N MARINE ENVIRONMENT WU: DN288604 6ý AUTHOR(S) Accesion For I...measure the acute and sublethal effects of heavy metals ( tributyltin , copper, and zinc) and storm drain effluent on the light output from marine...Grovhoug 3 THE USE OF STIM1ULABLE BIOLUMINESCENCE FROM MARINE DINOFLAGELLATES AS A MEANS OF DETECTING TOXICITY IN THE MARINE ENVIRONMENT. REFERENCE

Freshwater savings from marine protein consumption

International Nuclear Information System (INIS)

Gephart, Jessica A; Pace, Michael L; D’Odorico, Paolo

2014-01-01

Marine fisheries provide an essential source of protein for many people around the world. Unlike alternative terrestrial sources of protein, marine fish production requires little to no freshwater inputs. Consuming marine fish protein instead of terrestrial protein therefore represents freshwater savings (equivalent to an avoided water cost) and contributes to a low water footprint diet. These water savings are realized by the producers of alternative protein sources, rather than the consumers of marine protein. This study quantifies freshwater savings from marine fish consumption around the world by estimating the water footprint of replacing marine fish with terrestrial protein based on current consumption patterns. An estimated 7 600 km 3  yr −1 of water is used for human food production. Replacing marine protein with terrestrial protein would require an additional 350 km 3  yr −1 of water, meaning that marine protein provides current water savings of 4.6%. The importance of these freshwater savings is highly uneven around the globe, with savings ranging from as little as 0 to as much as 50%. The largest savings as a per cent of current water footprints occur in Asia, Oceania, and several coastal African nations. The greatest national water savings from marine fish protein occur in Southeast Asia and the United States. As the human population increases, future water savings from marine fish consumption will be increasingly important to food and water security and depend on sustainable harvest of capture fisheries and low water footprint growth of marine aquaculture. (paper)

United States of America. Report 1 [Marine Radioecology. Current Research and Future Scope

International Nuclear Information System (INIS)

Nakatani, R.E.

1967-01-01

Biology Department's aquatic research activities are primarily in freshwater radioecology and not in marine radioecology. The major reason for this lack of marine work is geographical location of our laboratory on the banks of the Columbia River, roughly 300 miles from the ocean. Recently, however, we have completed some preliminary work with 65 Zn metabolism in Anonyx sp., a marine benthic amphipod. Lake some inland laboratories, we have found that limited marine work can be done by hauling in sea water. We have been able to maintain the amphipods in a reasonably healthy state in our laboratory for about four weeks

Importance of benthonic marine flora monitoring in the liquid effluent discharge form Angra-1 Nuclear Power Plant

International Nuclear Information System (INIS)

Bloise, G.C.; Araujo Costa, D. de

1994-01-01

Angra-1 Nuclear Power Plant use sea water to condenser the steam of the secondary circuit. This water capted from Itaorna bay is chlorined and discharged more heater in Piraquara de Fora small bay. The temperature, chlorinade concentration, marine flora and fauna are monitored frequently with the intend of value the impact caused by this discharge to marine environment. The macroscopic marines algae is very sensible to environment temperature variations, constitutes on of the main rink in the food chain and stay every time attach at the bottom. Because of this facts they are considered an important bio indicators. (author). 5 refs, 2 figs, 2 tabs

Genetic Characterization of the Carotenoid Biosynthetic Pathway in Methylobacterium extorquens AM1 and Isolation of a Colorless Mutant

OpenAIRE

Van Dien, Stephen J.; Marx, Christopher J.; O'Brien, Brooke N.; Lidstrom, Mary E.

2003-01-01

Genomic searches were used to reconstruct the putative carotenoid biosynthesis pathway in the pink-pigmented facultative methylotroph Methylobacterium extorquens AM1. Four genes for putative phytoene desaturases were identified. A colorless mutant was obtained by transposon mutagenesis, and the insertion was shown to be in one of the putative phytoene desaturase genes. Mutations in the other three did not affect color. The tetracycline marker was removed from the original transposon mutant, r...

Otters, Marine

Science.gov (United States)

Estes, James A.; Bodkin, James L.; Ben-David, M.; Perrin, William F.; Würsing, Bernd; Thewissen, J.G.M.

2009-01-01

The otters (Mustelidae; Lutrinae) provide an exceptional perspective into the evolution of marine living by mammals. Most extant marine mammals (e.g. the cetaceans, pinnipeds, and sirenians) have been so highly modified by long periods of selection for life in the sea that they bear little resemblance to their terrestrial ancestors. Marine otters, in contrast, are more recent expatriates from freshwater habitats and some species still live in both environments. Contrasts among species within the otters, and among the otters, terrestrial mammals, and the more highly adapted pinnipeds and cetaceans provide powerful insights into mammalian adaptations to life in the sea (Estes, 1989). Among the marine mammals, sea otters (Enhydra lutris, Fig. 1) provide the clearest understanding of consumer-induced effects on ecosystem function. This is due in part to opportunities provided by history and in part to the relative ease with which shallow coastal systems where sea otters live can be observed and studied. Although more difficult to study than sea otters, other otter species reveal the connectivity among the marine, freshwater, and terrestrial systems. These three qualities of the otters – their comparative biology, their role as predators, and their role as agents of ecosystem connectivity – are what make them interesting to marine mammalogy.The following account provides a broad overview of the comparative biology and ecology of the otters, with particular emphasis on those species or populations that live in the sea. Sea otters are features prominently, in part because they live exclusively in the sea whereas other otters have obligate associations with freshwater and terrestrial environments (Kenyon, 1969; Riedman and Estes, 1990).

Developmental toxicity and molecular responses of marine medaka (Oryzias melastigma) embryos to ciguatoxin P-CTX-1 exposure.

Science.gov (United States)

Yan, Meng; Leung, Priscilla T Y; Ip, Jack C H; Cheng, Jin-Ping; Wu, Jia-Jun; Gu, Jia-Rui; Lam, Paul K S

2017-04-01

Ciguatoxins are produced by toxic benthic dinoflagellates and cause ciguatera fish poisoning worldwide, but the toxic effects on developing marine fish have not been well investigated. The Pacific ciguatoxin (P-CTX-1), is a potent sodium channel agonist, which is one of the most toxic members among all CTXs. This study evaluated the toxic effects of microinjecting purified Pacific ciguatoxin-1 (P-CTX-1) on embryonic development of marine medaka Oryzias melastigma. A lower 96h-LD 50 value was estimated for eleuthero-embryos (1.32ngg -1 ) than that for embryos (1.71ngg -1 ), indicating that P-CTX-1 is more lethal to newly hatched medaka larvae. P-CTX-1 induced detrimental effects during embryonic development, including hatching failure, abnormalities in physical development (caudal fin malformation and spinal deformities), internal damage (green coloration of the gall bladder and hemorrhaging), immune dysfunction, and altered muscle physiology (bradycardia and hyperkinetic twitching). The results of a transcriptional expression analysis of genes related to the stress/immune responses, cardiac and bone development, and apoptosis supported the observed developmental abnormalities. This study advanced the understanding of P-CTX-1 mediated toxic mechanisms in the development of early life stages of a fish, and thus contributed to the toxicity assessment of CTXs in marine ecosystems. Copyright © 2017 Elsevier B.V. All rights reserved.

Characterization of a marine-isolated mercury-resistant Pseudomonas putida strain SP1 and its potential application in marine mercury reduction

Energy Technology Data Exchange (ETDEWEB)

Zhang, Weiwei; Chen, Lingxin; Liu, Dongyan [Chinese Academy of Sciences, Yantai, SD (China). Yantai Inst. of Coastal Zone Research (YICCAS); Chinese Academy of Sciences, Yantai, SD (China). Shandong Provincial Key Lab. of Coastal Zone Environmental Processes

2012-02-15

The Pseudomonas putida strain SP1 was isolated from marine environment and was found to be resistant to 280 {mu}M HgCl{sub 2}. SP1 was also highly resistant to other metals, including CdCl{sub 2}, CoCl{sub 2}, CrCl{sub 3}, CuCl{sub 2}, PbCl{sub 2}, and ZnSO{sub 4}, and the antibiotics ampicillin (Ap), kanamycin (Kn), chloramphenicol (Cm), and tetracycline (Tc). mer operon, possessed by most mercury-resistant bacteria, and other diverse types of resistant determinants were all located on the bacterial chromosome. Cold vapor atomic absorption spectrometry and a volatilization test indicated that the isolated P. putida SP1 was able to volatilize almost 100% of the total mercury it was exposed to and could potentially be used for bioremediation in marine environments. The optimal pH for the growth of P. putida SP1 in the presence of HgCl{sub 2} and the removal of HgCl{sub 2} by P. putida SP1 was between 8.0 and 9.0, whereas the optimal pH for the expression of merA, the mercuric reductase enzyme in mer operon that reduces reactive Hg{sup 2+} to volatile and relatively inert monoatomic Hg{sup 0} vapor, was around 5.0. LD50 of P. putida SP1 to flounder and turbot was 1.5 x 10{sup 9} CFU. Biofilm developed by P. putida SP1 was 1- to 3-fold lower than biofilm developed by an aquatic pathogen Pseudomonas fluorescens TSS. The results of this study indicate that P. putida SP1 is a low virulence strain that can potentially be applied in the bioremediation of HgCl{sub 2} contamination over a broad range of pH. (orig.)

Sleeping Beauty transposon mutagenesis identifies genes that cooperate with mutant Smad4 in gastric cancer development.

Science.gov (United States)

Takeda, Haruna; Rust, Alistair G; Ward, Jerrold M; Yew, Christopher Chin Kuan; Jenkins, Nancy A; Copeland, Neal G

2016-04-05

Mutations in SMAD4 predispose to the development of gastrointestinal cancer, which is the third leading cause of cancer-related deaths. To identify genes driving gastric cancer (GC) development, we performed a Sleeping Beauty (SB) transposon mutagenesis screen in the stomach of Smad4(+/-) mutant mice. This screen identified 59 candidate GC trunk drivers and a much larger number of candidate GC progression genes. Strikingly, 22 SB-identified trunk drivers are known or candidate cancer genes, whereas four SB-identified trunk drivers, including PTEN, SMAD4, RNF43, and NF1, are known human GC trunk drivers. Similar to human GC, pathway analyses identified WNT, TGF-β, and PI3K-PTEN signaling, ubiquitin-mediated proteolysis, adherens junctions, and RNA degradation in addition to genes involved in chromatin modification and organization as highly deregulated pathways in GC. Comparative oncogenomic filtering of the complete list of SB-identified genes showed that they are highly enriched for genes mutated in human GC and identified many candidate human GC genes. Finally, by comparing our complete list of SB-identified genes against the list of mutated genes identified in five large-scale human GC sequencing studies, we identified LDL receptor-related protein 1B (LRP1B) as a previously unidentified human candidate GC tumor suppressor gene. In LRP1B, 129 mutations were found in 462 human GC samples sequenced, and LRP1B is one of the top 10 most deleted genes identified in a panel of 3,312 human cancers. SB mutagenesis has, thus, helped to catalog the cooperative molecular mechanisms driving SMAD4-induced GC growth and discover genes with potential clinical importance in human GC.

Chick derived induced pluripotent stem cells by the poly-cistronic transposon with enhanced transcriptional activity.

Science.gov (United States)

Katayama, Masafumi; Hirayama, Takashi; Tani, Tetsuya; Nishimori, Katsuhiko; Onuma, Manabu; Fukuda, Tomokazu

2018-02-01

Induced pluripotent stem (iPS) cell technology lead terminally differentiated cells into the pluripotent stem cells through the expression of defined reprogramming factors. Although, iPS cells have been established in a number of mammalian species, including mouse, human, and monkey, studies on iPS cells in avian species are still very limited. To establish chick iPS cells, six factors were used within the poly-cistronic reprogramming vector (PB-R6F), containing M3O (MyoD derived transactivation domain fused with Oct3/4), Sox2, Klf4, c-Myc, Lin28, and Nanog. The PB-R6F derived iPS cells were alkaline-phosphatase and SSEA-1 positive, which are markers of pluripotency. Elevated levels of endogenous Oct3/4 and Nanog genes were detected in the established iPS cells, suggesting the activation of the FGF signaling pathway is critical for the pluripotent status. Histological analysis of teratoma revealed that the established chick iPS cells have differentiation ability into three-germ-layer derived tissues. This is the first report of establishment of avian derived iPS cells with a single poly-cistronic transposon based expression system. The establishment of avian derived iPS cells could contribute to the genetic conservation and modification of avian species. © 2017 Wiley Periodicals, Inc.

Marine bioacoustics and technology: The new world of marine acoustic ecology

Science.gov (United States)

Hastings, Mardi C.; Au, Whitlow W. L.

2012-11-01

Marine animals use sound for communication, navigation, predator avoidance, and prey detection. Thus the rise in acoustic energy associated with increasing human activity in the ocean has potential to impact the lives of marine animals. Thirty years ago marine bioacoustics primarily focused on evaluating effects of human-generated sound on hearing and behavior by testing captive animals and visually observing wild animals. Since that time rapidly changing electronic and computing technologies have yielded three tools that revolutionized how bioacousticians study marine animals. These tools are (1) portable systems for measuring electrophysiological auditory evoked potentials, (2) miniaturized tags equipped with positioning sensors and acoustic recording devices for continuous short-term acoustical observation rather than intermittent visual observation, and (3) passive acoustic monitoring (PAM) systems for remote long-term acoustic observations at specific locations. The beauty of these breakthroughs is their direct applicability to wild animals in natural habitats rather than only to animals held in captivity. Hearing capabilities of many wild species including polar bears, beaked whales, and reef fishes have now been assessed by measuring their auditory evoked potentials. Miniaturized acoustic tags temporarily attached to an animal to record its movements and acoustic environment have revealed the acoustic foraging behavior of sperm and beaked whales. Now tags are being adapted to fishes in effort to understand their behavior in the presence of noise. Moving and static PAM systems automatically detect and characterize biological and physical features of an ocean area without adding any acoustic energy to the environment. PAM is becoming a powerful technique for understanding and managing marine habitats. This paper will review the influence of these transformative tools on the knowledge base of marine bioacoustics and elucidation of relationships between marine

Durability of precast prestressed concrete piles in marine environment : reinforcement corrosion and mitigation - Part 1.

Science.gov (United States)

2011-06-01

Research conducted in Part 1 has verified that precast prestressed concrete piles in : Georgias marine environment are deteriorating. The concrete is subjected to sulfate and : biological attack and the prestressed and nonprestressed reinforcement...

Marine biology

International Nuclear Information System (INIS)

Thurman, H.V.; Webber, H.H.

1984-01-01

This book discusses both taxonomic and ecological topics on marine biology. Full coverage of marine organisms of all five kingdoms is provided, along with interesting and thorough discussion of all major marine habitats. Organization into six major parts allows flexibility. It also provides insight into important topics such as disposal of nuclear waste at sea, the idea that life began on the ocean floor, and how whales, krill, and people interact. A full-color photo chapter reviews questions, and exercises. The contents are: an overview marine biology: fundamental concepts/investigating life in the ocean; the physical ocean, the ocean floor, the nature of water, the nature and motion of ocean water; general ecology, conditions for life in the sea, biological productivity and energy transfer; marine organisms; monera, protista, mycota and metaphyta; the smaller marine animals, the large animals marine habitats, the intertidal zone/benthos of the continental shelf, the photic zone, the deep ocean, the ocean under stress, marine pollution, appendix a: the metric system and conversion factors/ appendix b: prefixes and suffixes/ appendix c: taxonomic classification of common marine organisms, and glossary, and index

Development of novel metabolite-responsive transcription factors via transposon-mediated protein fusion.

Science.gov (United States)

Younger, Andrew K D; Su, Peter Y; Shepard, Andrea J; Udani, Shreya V; Cybulski, Thaddeus R; Tyo, Keith E J; Leonard, Joshua N

2018-02-01

Naturally evolved metabolite-responsive biosensors enable applications in metabolic engineering, ranging from screening large genetic libraries to dynamically regulating biosynthetic pathways. However, there are many metabolites for which a natural biosensor does not exist. To address this need, we developed a general method for converting metabolite-binding proteins into metabolite-responsive transcription factors-Biosensor Engineering by Random Domain Insertion (BERDI). This approach takes advantage of an in vitro transposon insertion reaction to generate all possible insertions of a DNA-binding domain into a metabolite-binding protein, followed by fluorescence activated cell sorting to isolate functional biosensors. To develop and evaluate the BERDI method, we generated a library of candidate biosensors in which a zinc finger DNA-binding domain was inserted into maltose binding protein, which served as a model well-studied metabolite-binding protein. Library diversity was characterized by several methods, a selection scheme was deployed, and ultimately several distinct and functional maltose-responsive transcriptional biosensors were identified. We hypothesize that the BERDI method comprises a generalizable strategy that may ultimately be applied to convert a wide range of metabolite-binding proteins into novel biosensors for applications in metabolic engineering and synthetic biology. © The Author(s) 2018. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Will marine productivity wane?

Science.gov (United States)

Laufkötter, Charlotte; Gruber, Nicolas

2018-03-01

If marine algae are impaired severely by global climate change, the resulting reduction in marine primary production would strongly affect marine life and the ocean's biological pump that sequesters substantial amounts of atmospheric carbon dioxide in the ocean's interior. Most studies, including the latest generation of Earth system models, project only moderate global decreases in biological production until 2100 (1, 2), suggesting that these concerns are unwarranted. But on page 1139 of this issue, Moore et al. (3) show that this conclusion might be shortsighted and that there may be much larger long-term changes in ocean productivity than previously appreciated.

United Kingdom. Report 1 [Marine Radioecology. Current Research and Future Scope

International Nuclear Information System (INIS)

Templeton, W.L.

1967-01-01

Present research programme (long-term): There are five separate research projects in this Group, viz: A. Low-level irradiation studies on marine and fresh-water fish eggs. B. Accumulation and. metabolism of radionuclides by marine organisms. C. Physico-chemical states of radionuclides in sea water, D. Dispersion and transport of radionuclides in the coastal waters of Windscale. E. Mechanisms of the toxicity of zinc to fish. Details of each project are outlined

33 CFR 3.40-15 - Sector New Orleans Marine Inspection Zone and Captain of the Port Zone; Marine Safety Unit Morgan...

Science.gov (United States)

2010-07-01

... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Sector New Orleans Marine....40-15 Sector New Orleans Marine Inspection Zone and Captain of the Port Zone; Marine Safety Unit Morgan City. Sector New Orleans' office is located in New Orleans, LA. A subordinate unit, Marine Safety...

Genome analysis of Pseudoalteromonas flavipulchra JG1 reveals various survival advantages in marine environment.

Science.gov (United States)

Yu, Min; Tang, Kaihao; Liu, Jiwen; Shi, Xiaochong; Gulder, Tobias A M; Zhang, Xiao-Hua

2013-10-16

Competition between bacteria for habitat and resources is very common in the natural environment and is considered to be a selective force for survival. Many strains of the genus Pseudoalteromonas were confirmed to produce bioactive compounds that provide those advantages over their competitors. In our previous study, P. flavipulchra JG1 was found to synthesize a Pseudoalteromonas flavipulchra antibacterial Protein (PfaP) with L-amino acid oxidase activity and five small chemical compounds, which were the main competitive agents of the strain. In addition, the genome of this bacterium has been previously sequenced as Whole Genome Shotgun project (PMID: 22740664). In this study, more extensive genomic analysis was performed to identify specific genes or gene clusters which related to its competitive feature, and further experiments were carried out to confirm the physiological roles of these genes when competing with other microorganisms in marine environment. The antibacterial protein PfaP may also participate in the biosynthesis of 6-bromoindolyl-3-acetic acid, indicating a synergistic effect between the antibacterial macromolecule and small molecules. Chitinases and quorum quenching enzymes present in P. flavipulchra, which coincide with great chitinase and acyl homoserine lactones degrading activities of strain JG1, suggest other potential mechanisms contribute to antibacterial/antifungal activities. Moreover, movability and rapid response mechanisms to phosphorus starvation and other stresses, such as antibiotic, oxidative and heavy metal stress, enable JG1 to adapt to deleterious, fluctuating and oligotrophic marine environments. The genome of P. flavipulchra JG1 exhibits significant genetic advantages against other microorganisms, encoding antimicrobial agents as well as abilities to adapt to various adverse environments. Genes involved in synthesis of various antimicrobial substances enriches the antagonistic mechanisms of P. flavipulchra JG1 and affords

Genome-Wide Transposon Mutagenesis Indicates that Mycobacterium marinum Customizes Its Virulence Mechanisms for Survival and Replication in Different Hosts

KAUST Repository

Weerdenburg, Eveline M.

2015-02-17

The interaction of environmental bacteria with unicellular eukaryotes is generally considered a major driving force for the evolution of intracellular pathogens, allowing them to survive and replicate in phagocytic cells of vertebrate hosts. To test this hypothesis on a genome-wide level, we determined for the intracellular pathogen Mycobacterium marinum whether it uses conserved strategies to exploit host cells from both protozoan and vertebrate origin. Using transposon-directed insertion site sequencing (TraDIS), we determined differences in genetic requirements for survival and replication in phagocytic cells of organisms from different kingdoms. In line with the general hypothesis, we identified a number of general virulence mechanisms, including the type VII protein secretion system ESX-1, biosynthesis of polyketide lipids, and utilization of sterols. However, we were also able to show that M. marinum contains an even larger set of host-specific virulence determinants, including proteins involved in the modification of surface glycolipids and, surprisingly, the auxiliary proteins of the ESX-1 system. Several of these factors were in fact counterproductive in other hosts. Therefore, M. marinum contains different sets of virulence factors that are tailored for specific hosts. Our data imply that although amoebae could function as a training ground for intracellular pathogens, they do not fully prepare pathogens for crossing species barriers.

Events Calendar: Smithsonian Marine Ecosystems Exhibit: Smithsonian Marine

Science.gov (United States)

current Smithsonian research on the plants and animals of the Indian River Lagoon and marine environments Station (SMS) at Fort Pierce Smithsonian Marine Station at Fort Pierce Website Search Box History Modeling Ecosystems Virtual Tour Facebook Instagram Twitter SMS Home › Smithsonian Marine

Polonium (210Po) and lead (210Pb) in marine organisms and their transfer in marine food chains

International Nuclear Information System (INIS)

Carvalho, Fernando P.

2011-01-01

The determination of 210 Po and 210 Pb was performed in marine organisms from the seashore to abyssal depths, encompassing a plethora of species from the microscopic plankton to the sperm whale. Concentrations of those radionuclides ranged from low values of about 5 x 10 -1 Bq kg -1 (wet wt.) in jellyfish, to very high values of about of 3 x 10 4 Bq kg -1 (wet wt.) in the gut walls of sardines, with a common pattern of 210 Po > 210 Pb.These radionuclides are primarily absorbed from water and concentrated by phyto- and microzooplankton, and then are transferred to the next trophic level along marine food chains. Investigation in epipelagic, mesopelagic, bathypelagic and abyssobenthic organisms revealed that 210 Po is transferred in the marine food webs with transfer factors ranging from 0.1 to 0.7, and numerically similar to those of the energy transfer in the marine food chains. As 210 Po preferentially binds to amino acids and proteins, its transfer in food chains likely traces protein transfer and, thus, 210 Po transfer factors are similar to ecotrophic coefficients. 210 Pb is transferred less efficiently in marine food chains and this contributes to increased 210 Po: 210 Pb activity ratios in some trophic levels.

Marine genomics

DEFF Research Database (Denmark)

Oliveira Ribeiro, Ângela Maria; Foote, Andrew David; Kupczok, Anne

2017-01-01

Marine ecosystems occupy 71% of the surface of our planet, yet we know little about their diversity. Although the inventory of species is continually increasing, as registered by the Census of Marine Life program, only about 10% of the estimated two million marine species are known. This lag......-throughput sequencing approaches have been helping to improve our knowledge of marine biodiversity, from the rich microbial biota that forms the base of the tree of life to a wealth of plant and animal species. In this review, we present an overview of the applications of genomics to the study of marine life, from...

Assessment of the toxicity of the solid coating PV1 in a marine invironment, using biotests with algae, a rotifer and a bacteria

NARCIS (Netherlands)

Foekema, E.M.; Sneekes, A.C.

2007-01-01

The toxic potential of substances that may leach from the solid coating PV1 was tested using • the marine bacteria Vibrio fisheri in the Microtox® Basic test • the marine algae Skeletonema costatum in a 72h algal growth inhibition test • the marine rotifer Brachionus plicatilis in the 24 h ROTOX®

Metabolites from marine fungus Aspergillus sp.

Digital Repository Service at National Institute of Oceanography (India)

Wahidullah, S.; Rajmanickam, R.; DeSouza, L.

Chemical examination of a methanolic extract of the marine fungus, Aspergillus sp., isolated from marine grass environment, yielded a steroid, ergosterol peroxide (1), and a mixture of known glyceride esters (2,3) of unsaturated fatty acids...

Marine environment news Vol. 4, no. 1, June 2006

International Nuclear Information System (INIS)

2006-06-01

The last six months have been a frenetically busy time for us in Monaco. Our Marine Programmes have been positively reviewed by the Standing Advisory Group on Nuclear Applications (SAGNA) and by an External Evaluation of our Programme. Both Groups report to the Director General, Mr Mohammed ElBaradei, and we hope that new investment in personnel and equipment may eventually result from their evaluations and feedback. We were honoured by the visit of His Serene Highness Prince Albert II of Monaco in March 2006 to our facilities. HSH continues to take a personal interest in MEL's isotopic and pollutant analyses of biota and environmental samples from the Arctic environment which we sampled during His Highness' cruise in June 2005 (see Vol. 3. No 2. MEL Newsletter). This issue also shows that MEL has hosted several important workshops and meetings. The US Research Vessel Endeavour visited the port of Monaco in April and MEL hosted an informal reception for the crew. The visit was in connection with ongoing, joint MEL-US studies in ocean carbon sinks in the Mediterranean (the MEDFLUX programme). More recently, MEL has been involved in discussion with Gulf Member States for a Marine Radioactivity Baseline Study. Finally, I am pleased to note that our MEL Newsletter is clearly having a positive outreach with Member States, since we are currently witnessing a doubling in Member States requests through the TC Concept Proposals (2007-2008) for fellowships, courses and capacity building in marine environment

Global patterns of extinction risk in marine and non-marine systems.

Science.gov (United States)

Webb, Thomas J; Mindel, Beth L

2015-02-16

Despite increasing concern over the effects of human activities on marine ecosystems, extinction in the sea remains scarce: 19-24 out of a total of >850 recorded extinctions implies a 9-fold lower marine extinction rate compared to non-marine systems. The extent of threats faced by marine systems, and their resilience to them, receive considerable attention, but the detectability of marine extinctions is less well understood. Before its extinction or threat status is recorded, a species must be both taxonomically described and then formally assessed; lower rates of either process for marine species could thus impact patterns of extinction risk, especially as species missing from taxonomic inventories may often be more vulnerable than described species. We combine data on taxonomic description with conservation assessments from the International Union for Conservation of Nature (IUCN) to test these possibilities across almost all marine and non-marine eukaryotes. We find that the 9-fold lower rate of recorded extinctions and 4-fold lower rate of ongoing extinction risk across marine species can be explained in part by differences in the proportion of species assessed by the IUCN (3% cf. 4% of non-marine species). Furthermore, once taxonomic knowledge and conservation assessments pass a threshold level, differences in extinction risk between marine and non-marine groups largely disappear. Indeed, across the best-studied taxonomic groups, there is no difference between marine and non-marine systems, with on average between 20% and 25% of species being threatened with extinction, regardless of realm. Copyright © 2015 Elsevier Ltd. All rights reserved.

Marine ecology

International Nuclear Information System (INIS)

Anon.

1977-01-01

Studies on marine ecology included marine pollution; distribution patterns of Pu and Am in the marine waters, sediments, and organisms of Bikini Atoll and the influence of physical, chemical, and biological factors on their movements through marine biogeochemical systems; transfer and dispersion of organic pollutants from an oil refinery through coastal waters; transfer of particulate pollutants, including sediments dispersed during construction of offshore power plants; and raft culture of the mangrove oysters

Marine Pollution and Ecotoxicology

Digital Repository Service at National Institute of Oceanography (India)

Sarkar, A.

. Heavy metals Editorial Guest The special issue of Environment International has come up with selected papers presented in the International workshop on Marine Pollution and Ecotoxicology held at the National Institute of Oceanography, Dona Paula, Goa... presented in this special issue are classified into five sections namely, Coastal water quality, Heavy metals, Trace metals, Persistent organic pollutants and Ecotoxicology. 1. Coastal water quality assessment The pollution of the marine environment has...

[Annotation of the mobilomes of nine teleost species].

Science.gov (United States)

Gao, Bo; Shen, Dan; Chen, Cai; Wang, Saisai; Yang, Kunlun; Chen, Wei; Wang, Wei; Zhang, Li; Song, Chengyi

2018-01-25

In this study, the mobilomes of nine teleost species were annotated by bioinformatics methods. Both of the mobilome size and constitute displayed a significant difference in 9 species of teleost fishes. The species of mobilome content ranking from high to low were zebrafish, medaka, tilapia, coelacanth, platyfish, cod, stickleback, tetradon and fugu. Mobilome content and genome size were positively correlated. The DNA transposons displayed higher diversity and larger variation in teleost (0.50% to 38.37%), was a major determinant of differences in teleost mobilomes, and hAT and Tc/Mariner superfamily were the major DNA transposons in teleost. RNA transposons also exhibited high diversity in teleost, LINE transposons accounted for 0.53% to 5.75% teleost genomic sequences, and 14 superfamilies were detected. L1, L2, RTE and Rex retrotransposons obtained significant amplification. While LTR displayed low amplification in most teleost with less than 2% of genome coverages, except in zebrafish and stickleback, where LTR reachs 5.58% and 2.51% of genome coverages respectively. And 6 LTR superfamilies (Copia, DIRS, ERV, Gypsy, Ngaro and Pao) were detected in the teleost, and Gypsy exhibits obvious amplication among them. While the SINE represents the weakest ampification types in teleost, only within zebrafish and coelacanth, it represents 3.28% and 5.64% of genome coverages, in the other 7 teleost, it occupies less than 1% of genomes, and tRNA, 5S and MIR families of SINE have a certain degree of amplification in some teleosts. This study shows that the teleost display high diversity and large variation of mobilome, there is a strong correlation with the size variations of genomes and mobilome contents in teleost, mobilome is an important factor in determining the teleost genome size.

Polonium (210Po) and lead (210Pb) in marine organisms and their transfer in marine food chains.

Science.gov (United States)

Carvalho, Fernando P

2011-05-01

The determination of (210)Po and (210)Pb was performed in marine organisms from the seashore to abyssal depths, encompassing a plethora of species from the microscopic plankton to the sperm whale. Concentrations of those radionuclides ranged from low values of about 5 × 10(-1) Bq kg(-1) (wet wt.) in jellyfish, to very high values of about of 3 × 10(4) Bq kg(-1) (wet wt.) in the gut walls of sardines, with a common pattern of (210)Po > (210)Pb.These radionuclides are primarily absorbed from water and concentrated by phyto- and microzooplankton, and then are transferred to the next trophic level along marine food chains. Investigation in epipelagic, mesopelagic, bathypelagic and abyssobenthic organisms revealed that (210)Po is transferred in the marine food webs with transfer factors ranging from 0.1 to 0.7, and numerically similar to those of the energy transfer in the marine food chains. As (210)Po preferentially binds to amino acids and proteins, its transfer in food chains likely traces protein transfer and, thus, (210)Po transfer factors are similar to ecotrophic coefficients. (210)Pb is transferred less efficiently in marine food chains and this contributes to increased (210)Po:(210)Pb activity ratios in some trophic levels. Copyright © 2010 Elsevier Ltd. All rights reserved.

33 CFR 3.40-28 - Sector Houston-Galveston Marine Inspection Zone and Captain of the Port Zone; Marine Safety Unit...

Science.gov (United States)

2010-07-01

... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Sector Houston-Galveston Marine....40-28 Sector Houston-Galveston Marine Inspection Zone and Captain of the Port Zone; Marine Safety Unit Port Arthur. Sector Houston-Galveston's office is located in Galena Park, TX. A subordinate unit...

Marine Viruses: Key Players in Marine Ecosystems

Directory of Open Access Journals (Sweden)

Mathias Middelboe

2017-10-01

Full Text Available Viruses were recognized as the causative agents of fish diseases, such as infectious pancreatic necrosis and Oregon sockeye disease, in the early 1960s [1], and have since been shown to be responsible for diseases in all marine life from bacteria to protists, mollusks, crustaceans, fish and mammals [2].[...

Mitochondrial Cytochrome oxidase 1 phylogeny supports alternative taxonomic scheme for the marine Haplosclerida

NARCIS (Netherlands)

Raleigh, J.; Redmond, N.E.; Delahan, E.; Tropey, S.; van Soest, R.W.M.; Kelly, M.; McCormack, G.P.

2007-01-01

Recent molecular studies have shown that the sponge order Haploslcerida is polyphyletic as the freshwater sponges appear to be more closely related to other demosponges than they are to the marine haplosclerids. Within the marine haplosclerid clade relationships viewed via 18S and 28S rRNA gene

46 CFR 35.20-1 - Notice to mariners; aids to navigation-T/OCLB.

Science.gov (United States)

2010-10-01

... Notices to Mariners (Great Lakes Edition), published by the Commander, 9th Coast Guard District, contain... of navigation on the Great Lakes. These notices may be obtained free of charge, by making application to Commander, 9th Coast Guard District. (c) Weekly Notices to Mariners (worldwide coverage) are...

Marine Science Teaching at the University Level. Report of the Unesco Workshop on University Curricula. Unesco Technical Papers in Marine Science No. 19.

Science.gov (United States)

United Nations Educational, Scientific, and Cultural Organization, Paris (France). Div. of Marine Sciences.

A group of marine science education educators from several countries were requested to provide guidelines for the education and training of marine scientists and formulate recommended curricula in the following disciplines: marine biology (including fisheries biology), physical oceanography, and marine geology. Included in the report are: (1)…

26 CFR 1.831-1 - Tax on insurance companies (other than life or mutual), mutual marine insurance companies, and...

Science.gov (United States)

2010-04-01

... mutual), mutual marine insurance companies, and mutual fire insurance companies issuing perpetual... companies (other than life or mutual), mutual marine insurance companies, and mutual fire insurance... insurance companies and mutual fire insurance companies exclusively issuing either perpetual policies, or...

Marine pollution

International Nuclear Information System (INIS)

Albaiges, J.

1989-01-01

This book covers the following topics: Transport of marine pollutants; Transformation of pollutants in the marine environment; Biological effects of marine pollutants; Sources and transport of oil pollutants in the Persian Gulf; Trace metals and hydrocarbons in Syrian coastal waters; and Techniques for analysis of trace pollutants

Genetic Characterization of the Carotenoid Biosynthetic Pathway in Methylobacterium extorquens AM1 and Isolation of a Colorless Mutant

Science.gov (United States)

Van Dien, Stephen J.; Marx, Christopher J.; O'Brien, Brooke N.; Lidstrom, Mary E.

2003-01-01

Genomic searches were used to reconstruct the putative carotenoid biosynthesis pathway in the pink-pigmented facultative methylotroph Methylobacterium extorquens AM1. Four genes for putative phytoene desaturases were identified. A colorless mutant was obtained by transposon mutagenesis, and the insertion was shown to be in one of the putative phytoene desaturase genes. Mutations in the other three did not affect color. The tetracycline marker was removed from the original transposon mutant, resulting in a pigment-free strain with wild-type growth properties useful as a tool for future experiments. PMID:14660416

Genetic characterization of the carotenoid biosynthetic pathway in Methylobacterium extorquens AM1 and isolation of a colorless mutant.

Science.gov (United States)

Van Dien, Stephen J; Marx, Christopher J; O'Brien, Brooke N; Lidstrom, Mary E

2003-12-01

Genomic searches were used to reconstruct the putative carotenoid biosynthesis pathway in the pink-pigmented facultative methylotroph Methylobacterium extorquens AM1. Four genes for putative phytoene desaturases were identified. A colorless mutant was obtained by transposon mutagenesis, and the insertion was shown to be in one of the putative phytoene desaturase genes. Mutations in the other three did not affect color. The tetracycline marker was removed from the original transposon mutant, resulting in a pigment-free strain with wild-type growth properties useful as a tool for future experiments.

Marine Corps Expeditionary Rifle Platoon Energy Burden

Science.gov (United States)

2014-12-01

26  a.  Step 1: The Flight to MRP AOR ............................................26  b...Terrain and Troops MEU Marine Expeditionary Unit MRP Marine Rifle Platoon NVG Night Vision Goggles SAW Squad automatic weapon SPOD Seaport...rifle platoon ( MRP ). The MRP is an infantry unit, which is the core component of the GCE. Each MRP consists of 40–45 Marines and requires energy

Implications of S1 tephra findings in Dead Sea and Tayma palaeolake sediments for marine reservoir age estimation and palaeoclimate synchronisation

Science.gov (United States)

Neugebauer, Ina; Wulf, Sabine; Schwab, Markus J.; Serb, Johanna; Plessen, Birgit; Appelt, Oona; Brauer, Achim

2017-08-01

Here we report on the first findings of a cryptotephra in the Holocene lacustrine sediment records of the Dead Sea and Tayma palaeolake (NW Arabian Peninsula). The major element glass composition of this rhyolitic tephra is identical to the distal 'S1' tephra layer identified in the Yammoûneh palaeolake (Lebanon), in a marine sediment record from the SE Levantine basin and in the Sodmein Cave archaeological site in Egypt. The S1 tephra corresponds to the early Holocene 'Dikkartın' dome eruption of the Erciyes Dağ volcano in central Anatolia (Turkey) and has been dated in the marine record at 8830 ± 140 cal yr BP. We present new age estimates of the S1 tephra based on radiocarbon dating of terrestrial plant remains and pollen concentrates revealing ages of 8939 ± 83 cal yr BP in the Dead Sea sediments and 9041 ± 254 cal yr BP in Tayma. The precise date from the Dead Sea allows refining the early Holocene marine reservoir age in the SE Levantine Sea to ca. 320 ± 50 years. Synchronisation of marine and terrestrial palaeoclimate records in the eastern Mediterranean region using the S1 tephra further suggests a time-transgressive expansion of the early Holocene humid period.

Ecotoxicology of arsenic in the marine environment

Energy Technology Data Exchange (ETDEWEB)

Neff, J.M. [Battelle Ocean Sciences Lab., Duxbury, MA (United States)

1997-05-01

Arsenic has a complex marine biogeochemistry that has important implications for its toxicity to marine organisms and their consumers. The average concentration of total arsenic in the ocean is about 1.7 {micro}g/L, about two orders of magnitude higher than the US Environmental Protection Agency`s human health criterion value of 0.0175 {micro}g/L. The dominant form of arsenic in oxygenated marine and brackish waters in arsenate (As V). The more toxic and potentially carcinogenic arsenite (As III) rarely accounts for more than 20% of total arsenic in seawater. Uncontaminated marine sediments contain from 5 to about 40 {micro}g/g dry weight total arsenic. Arsenate dominates in oxidized sediments and is associated primarily with iron oxyhydroxides. In reducing marine sediments, arsenate is reduced to arsenite and is associated primarily with sulfide minerals. Marine algae accumulate arsenate from seawater, reduce it to arsenite, and then oxidize the arsenite to a large number of organoarsenic compounds. The algae release arsenite, methylarsonic acid, and dimethylarsinic acid to seawater. Dissolved arsenite and arsenate are more toxic to marine phytoplankton than to marine invertebrates and fish. This may be due to the fact that marine animals have a limited ability to bioconcentrate inorganic arsenic from seawater but can bioaccumulate organoarsenic compounds from their food. Tissues of marine invertebrates and fish contain high concentrations of arsenic, usually in the range of about 1 to 100 {micro}g/g dry weight, most of it in the form of organoarsenic compounds, particularly arsenobetaine. Organoarsenic compounds are bioaccumulated by human consumers of seafood products, but the arsenic is excreted rapidly, mostly as organoarsenic compounds. Arsenobetaine, the most abundant organoarsenic compound in seafoods, is not toxic or carcinogenic to mammals. Little of the organoarsenic accumulated by humans from seafood is converted to toxic inorganic arsenite.

The Consolidation of Administrative Functions for U.S. Marine Forces, Pacific

National Research Council Canada - National Science Library

Allen, Patrick

2004-01-01

Marine Administrative Message 441/99 directed that Marine Corps administration be consolidated above the battalion level, Secondly, Marine Administrative Message 027/04 directed that over 1,300 Marine...

Proceedings of the 2008 marine biodiesel symposium

International Nuclear Information System (INIS)

2008-01-01

In addition to producing lower hydrocarbon emissions, marine biodiesel is biodegradable and does not harm fish. This symposium was held to discuss current marine biodiesel applications and examine methods of increasing the use of biodiesel in marine environments in British Columbia (BC). Biofuel policies and mandates in the province were reviewed, and methods of expanding the biodiesel market were explored. Updates on the use of biodiesel in ferries, tugboats, and smaller marine diesel engine applications were provided. Biodiesel projects in the United States were discussed. The environmental impacts of marine biodiesel were evaluated, and federal policies and standards for biodiesel were also outlined. The symposium was divided into the following 5 main sessions: (1) policy, (2) overviews, (3) using biodiesel in marine engines, (4) biodiesel in larger marine vessels, and (5) biodiesel quality and environmental considerations. The conference featured 13 presentations, of which 4 have been catalogued separately for inclusion in this database. tabs., figs

Collaboration or Contention? Decentralised marine Governance in Berau

NARCIS (Netherlands)

Kusumawati, R.; Visser, L.E.

2014-01-01

Conservation of marine space is a new frontier in environmentalists’ involvement with resource governance in Indonesia. The coastal and marine area of Berau was established as a District Marine Conservation Area (MCA) based on District Head Regulation No. 31/2005. The total MCA of 1.27 million ha

New Waves in Marine Science Symposium: Marine Animal Communication.

Science.gov (United States)

Allen, Betty, Comp.

1989-01-01

Presented are the abstracts from three research projects on marine social systems which were a part of a marine science symposium. Five sets of activities on marine animal communication are included, one each for grades K-2, 3-5, 6-8 and 9-12, and informal education. (CW)

Marine radioecology. Final reports from sub-projects within the Nordic nuclear safety research project EKO-1

International Nuclear Information System (INIS)

Palsson, S.E.

2001-04-01

This report contains a collection of eight papers describing research done in the NKS/EKO-1 project. It also contains a preface giving a summary of the results. The EKO-1 project as a whole has been described in the report NKS(97)FR4. The aim of the project was to make a joint Nordic study on radionuclides in sediments and water and the interaction between these two phaseS. Relatively less emphasis had been put on this factor compared to others in previous Nordic studies on marine radioecology. For some of the participating countries this work was the first of its kind undertaken. The project involved field, laboratory and model studies. The work and results helped to highlight the important role of sediments when assessing the consequences of real or possible releases of radionuclides to the marine environment (au)

Insertion of transposon in the vicinity of SSK2 confers enhanced tolerance to furfural in Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Kim, Hyun-Soo [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Science; Kim, Na-Rae [Ewha Womans Univ., Seoul (Korea, Republic of). Div. of Life and Pharmaceutical Sciences; Kim, Wankee [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Science; Ajou Univ., Suwon (Korea, Republic of). Inst. for Medical Sciences; Choi, Wonja [Ewha Womans Univ., Seoul (Korea, Republic of). Dept. of Life Science; Ewha Womans Univ., Seoul (Korea, Republic of). Microbial Resources Research Center

2012-07-15

Furfural is one of the major inhibitors generated during sugar production from cellulosic materials and, as an aldehyde, inhibits various cellular activities of microorganisms used, leading to prolonged lag time during ethanologenic fermentation. Since Saccharomyces cerevisiae strains tolerant to furfural are of great economic benefit in producing bioethanol, much effort to obtain more efficient strains continues to be made. In this study, we examined the furfural tolerance of transposon mutant strains (Tn 1-5) with enhanced ethanol tolerance and found that one of them (Tn 2), in which SSK2 is downregulated at the transcriptional level, displayed improved furfural tolerance. Such phenotype was abolished by complementation of the entire open reading frame of SSK2, which encodes a mitogen-activated protein (MAP) kinase kinase kinase of the high osmolarity glycerol (HOG) signaling pathway, suggesting an inhibitory effect of SSK2 in coping with furfural stress. Tn 2 showed a significant decrease in the intracellular level of reactive oxygen species (ROS) and early and high activation of Hog1p, a MAP kinase integral to the HOG pathway in response to furfural. The transcriptional levels of CTT1 and GLR1, two of known Hog1p downstream target genes whose protein products are involved in reducing ROS, were increased by 43 % and 56 % respectively compared with a control strain, probably resulting in the ROS decrease. Tn 2 also showed a shortened lag time during fermentation in the presence of furfural, resulting from efficient conversion of furfural to non-toxic (or less toxic) furfuryl alcohol. Taken together, the enhanced furfural tolerance of Tn 2 is suggested to be conferred by the combined effect of an early event of less ROS accumulation and a late event of efficient detoxification of furfural. (orig.)

Insertion of transposon in the vicinity of SSK2 confers enhanced tolerance to furfural in Saccharomyces cerevisiae.

Science.gov (United States)

Kim, Hyun-Soo; Kim, Na-Rae; Kim, Wankee; Choi, Wonja

2012-07-01

Furfural is one of the major inhibitors generated during sugar production from cellulosic materials and, as an aldehyde, inhibits various cellular activities of microorganisms used, leading to prolonged lag time during ethanologenic fermentation. Since Saccharomyces cerevisiae strains tolerant to furfural are of great economic benefit in producing bioethanol, much effort to obtain more efficient strains continues to be made. In this study, we examined the furfural tolerance of transposon mutant strains (Tn 1-5) with enhanced ethanol tolerance and found that one of them (Tn 2), in which SSK2 is downregulated at the transcriptional level, displayed improved furfural tolerance. Such phenotype was abolished by complementation of the entire open reading frame of SSK2, which encodes a mitogen-activated protein (MAP) kinase kinase kinase of the high osmolarity glycerol (HOG) signaling pathway, suggesting an inhibitory effect of SSK2 in coping with furfural stress. Tn 2 showed a significant decrease in the intracellular level of reactive oxygen species (ROS) and early and high activation of Hog1p, a MAP kinase integral to the HOG pathway in response to furfural. The transcriptional levels of CTT1 and GLR1, two of known Hog1p downstream target genes whose protein products are involved in reducing ROS, were increased by 43 % and 56 % respectively compared with a control strain, probably resulting in the ROS decrease. Tn 2 also showed a shortened lag time during fermentation in the presence of furfural, resulting from efficient conversion of furfural to non-toxic (or less toxic) furfuryl alcohol. Taken together, the enhanced furfural tolerance of Tn 2 is suggested to be conferred by the combined effect of an early event of less ROS accumulation and a late event of efficient detoxification of furfural.

Marine Mammals and Noise-Progress Since 1995

Science.gov (United States)

2015-09-30

1 DISTRIBUTION STATEMENT A. Approved for public release; distribution is unlimited. Marine Mammals & Noise - Progress Since 1995 Christine...effects of underwater noise on marine mammals has grown steadily over the last few decades. Results and information are scattered across the peer...reviewed and grey literature. In this project, we try to review the information on noise impacts on marine mammals , focussing on results since the

High abundance of JS-1- and Chloroflexi-related Bacteria in deeply buried marine sediments revealed by quantitative, real-time PCR.

Science.gov (United States)

Blazejak, Anna; Schippers, Axel

2010-05-01

Sequences of members of the bacterial candidate division JS-1 and the classes Anaerolineae and Caldilineae of the phylum Chloroflexi are frequently found in 16S rRNA gene clone libraries obtained from marine sediments. Using a newly designed quantitative, real-time PCR assay, these bacterial groups were jointly quantified in samples from near-surface and deeply buried marine sediments from the Peru margin, the Black Sea, and a forearc basin off the island of Sumatra. In near-surface sediments, sequences of the JS-1 as well as Anaerolineae- and Caldilineae-related Bacteria were quantified with significantly lower 16S rRNA gene copy numbers than the sequences of total Bacteria. In contrast, in deeply buried sediments below approximately 1 m depth, similar quantities of the 16S rRNA gene copies of these specific groups and Bacteria were found. This finding indicates that JS-1 and Anaerolineae- and Caldilineae-related Bacteria might dominate the bacterial community in deeply buried marine sediments and thus seem to play an important ecological role in the deep biosphere.

Marine Robot Autonomy

CERN Document Server

2013-01-01

Autonomy for Marine Robots provides a timely and insightful overview of intelligent autonomy in marine robots. A brief history of this emerging field is provided, along with a discussion of the challenges unique to the underwater environment and their impact on the level of intelligent autonomy required.  Topics covered at length examine advanced frameworks, path-planning, fault tolerance, machine learning, and cooperation as relevant to marine robots that need intelligent autonomy.  This book also: Discusses and offers solutions for the unique challenges presented by more complex missions and the dynamic underwater environment when operating autonomous marine robots Includes case studies that demonstrate intelligent autonomy in marine robots to perform underwater simultaneous localization and mapping  Autonomy for Marine Robots is an ideal book for researchers and engineers interested in the field of marine robots.      

Opportunities in Marine and Maritime Careers.

Science.gov (United States)

Heitzmann, Wm. Ray

This book describes careers related to the sea. The following chapters are included: (1) "The World of Water"; (2) "Cruise Ship Careers"; (3) "Oceanography and the Marine Sciences"; (4) "Fishing"; (5) "Commerical Diving"; (6) "Maritime Transportation"; (7) "Shipbuilding"; (8) "Military Careers Afloat"; (9) "Miscellaneous Marine and Maritime…

2017 Marine Hydrokinetic Instrumentation Workshop Report

Energy Technology Data Exchange (ETDEWEB)

Driscoll, Frederick R [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Mauer, Erik [U.S. Department of Energy; Rieks, Jeff [Allegheny Science and Technologies

2018-03-06

The third Marine Hydrokinetic Instrumentation Workshop was held at Florida Atlantic University's Sea Tech Campus in Dania Beach, Florida, from February 28 to March 1, 2017. The workshop brought together 37 experts in marine energy measurement, testing, and technology development to present and discuss the instrumentation and data-processing needs of the marine energy industry. The workshop was comprised of a plenary session followed by two focused breakout sessions. The half-day plenary session reviewed findings from prior instrumentation workshops, presented research activities that aim to fill previously identified gaps, and had industry experts present the state of the marine energy measurement technologies. This report provides further detail on the workshop, objectives, and findings.

Expression of phage-transposons of Pseudomonas aeruginosa in cells of Pseudomonas putida PpGl. II. Zygotic induction, an essential condition for the emergence of defective lysogens

Energy Technology Data Exchange (ETDEWEB)

Gorbunova, S.A.; Akhverdyan, V.Z.; Krylov, V.N.

1986-06-01

The presence of a large number of clones, which have lost the ability to produce phase, is a feature of PpGl exconjugants, which have acquired markers of the hybrid plasmid containing the genome of the PAO1 phage transposon. Analysis of these clones indicates that they contain plasmids with different defects in the phage genome (point mutations and different deletion lengths which may have an effect on both the phage genome and also plasmid RP4). Mutations of a particular region are selected, the region of early prophage genes. The process of zygotic induction is an essential condition for the arisal of mutants (including deletion mutants). The results of the experiment, which was based on the Luria-Delbruik test, showed that a significant proportion of the mutations, including deletion mutations, arise in the recipient cells PpGl.

Marine habitat mapping at Labuan Marine Park, Federal Territory of Labuan, Malaysia

Science.gov (United States)

Mustajap, Fazliana; Saleh, Ejria; Madin, John; Hamid, Shahimah Abdul

2015-06-01

Marine habitat mapping has recently become essential in coastal marine science research. It is one of the efforts to understand marine ecosystems, and thus to protect them. Habitat mapping is integral to marine-related industries such as fisheries, aquaculture, forestry and tourism. An assessment of marine habitat mapping was conducted at Labuan Marine Park (LMP), a marine protected area in the Federal Territory of Labuan. It is surrounded by shallow water within its islands (Kuraman, Rusukan Kecil and Rusukan Besar) with an area of 39.7 km2. The objectives of the study are to identify the substrate and types of marine habitat present within the park. Side scan sonar (SSS) (Aquascan TM) was used to determine the substrates and habitat while ground truthings were done through field observation and SCUBA diving survey. Seabed classification and marine habitat was based on NOAA's biogeography program. Three substrate types (sand, rock, silt) were identified in this area. The major marine habitats identified are corals, macro algae and small patches of sea grass. The study area is an important refuge for spawning and juvenile fish and supports the livelihood of the coastal communities on Labuan Island. Therefore, proper management is crucial in order to better maintain the marine protected area. The findings are significant and provide detailed baseline information on marine habitat for conservation, protection and future management in LMP.

2011 Marine Hydrokinetic Device Modeling Workshop: Final Report; March 1, 2011

Energy Technology Data Exchange (ETDEWEB)

Li, Y.; Reed, M.; Smith, B.

2011-10-01

This report summarizes the NREL Marine and Hydrokinetic Device Modeling Workshop. The objectives for the modeling workshop were to: (1) Review the designs of existing MHK device prototypes and discuss design and optimization procedures; (2) Assess the utility and limitations of modeling techniques and methods presently used for modeling MHK devices; (3) Assess the utility and limitations of modeling methods used in other areas, such as naval architecture and ocean engineering (e.g., oil & gas industry); and (4) Identify the necessary steps to link modeling with other important components that analyze MHK devices (e.g., tank testing, PTO design, mechanical design).

Genomic differentiation among two strains of the PS1 clade isolated from geographically separated marine habitats

KAUST Repository

Jimenez Infante, Francy M.; Ngugi, David; Alam, Intikhab; Rashid, Mamoon; Ba Alawi, Wail; Kamau, Allan; Bajic, Vladimir B.; Stingl, Ulrich

2014-01-01

Using dilution-to-extinction cultivation, we isolated a strain affiliated with the PS1 clade from surface waters of the Red Sea. Strain RS24 represents the second isolate of this group of marine Alphaproteobacteria after IMCC14465 that was isolated

77 FR 32571 - Marine Mammals; File No. 14856

Science.gov (United States)

2012-06-01

... Bruce R. Mate, Ph.D., Hatfield Marine Science Center, Oregon State University, Newport, OR 97365, has applied in due form for a permit to take marine mammals world-wide for the purposes of scientific research... identified species of marine mammals species world-wide. The purposes of the proposed research are to: (1...

Phylogenomics of Rhodobacteraceae reveals evolutionary adaptation to marine and non-marine habitats.

Science.gov (United States)

Simon, Meinhard; Scheuner, Carmen; Meier-Kolthoff, Jan P; Brinkhoff, Thorsten; Wagner-Döbler, Irene; Ulbrich, Marcus; Klenk, Hans-Peter; Schomburg, Dietmar; Petersen, Jörn; Göker, Markus

2017-06-01

Marine Rhodobacteraceae (Alphaproteobacteria) are key players of biogeochemical cycling, comprise up to 30% of bacterial communities in pelagic environments and are often mutualists of eukaryotes. As 'Roseobacter clade', these 'roseobacters' are assumed to be monophyletic, but non-marine Rhodobacteraceae have not yet been included in phylogenomic analyses. Therefore, we analysed 106 genome sequences, particularly emphasizing gene sampling and its effect on phylogenetic stability, and investigated relationships between marine versus non-marine habitat, evolutionary origin and genomic adaptations. Our analyses, providing no unequivocal evidence for the monophyly of roseobacters, indicate several shifts between marine and non-marine habitats that occurred independently and were accompanied by characteristic changes in genomic content of orthologs, enzymes and metabolic pathways. Non-marine Rhodobacteraceae gained high-affinity transporters to cope with much lower sulphate concentrations and lost genes related to the reduced sodium chloride and organohalogen concentrations in their habitats. Marine Rhodobacteraceae gained genes required for fucoidan desulphonation and synthesis of the plant hormone indole 3-acetic acid and the compatible solutes ectoin and carnitin. However, neither plasmid composition, even though typical for the family, nor the degree of oligotrophy shows a systematic difference between marine and non-marine Rhodobacteraceae. We suggest the operational term 'Roseobacter group' for the marine Rhodobacteraceae strains.

Phenotypic and genotypic description of Sedimenticola selenatireducens strain CUZ, a marine (per)chlorate-respiring gammaproteobacterium, and its close relative the chlorate-respiring Sedimenticola strain NSS.

Science.gov (United States)

Carlström, Charlotte I; Loutey, Dana E; Wang, Ouwei; Engelbrektson, Anna; Clark, Iain; Lucas, Lauren N; Somasekhar, Pranav Y; Coates, John D

2015-04-01

Two (per)chlorate-reducing bacteria, strains CUZ and NSS, were isolated from marine sediments in Berkeley and San Diego, CA, respectively. Strain CUZ respired both perchlorate and chlorate [collectively designated (per)chlorate], while strain NSS respired only chlorate. Phylogenetic analysis classified both strains as close relatives of the gammaproteobacterium Sedimenticola selenatireducens. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) preparations showed the presence of rod-shaped, motile cells containing one polar flagellum. Optimum growth for strain CUZ was observed at 25 to 30 °C, pH 7, and 4% NaCl, while strain NSS grew optimally at 37 to 42 °C, pH 7.5 to 8, and 1.5 to 2.5% NaCl. Both strains oxidized hydrogen, sulfide, various organic acids, and aromatics, such as benzoate and phenylacetate, as electron donors coupled to oxygen, nitrate, and (per)chlorate or chlorate as electron acceptors. The draft genome of strain CUZ carried the requisite (per)chlorate reduction island (PRI) for (per)chlorate respiration, while that of strain NSS carried the composite chlorate reduction transposon responsible for chlorate metabolism. The PRI of strain CUZ encoded a perchlorate reductase (Pcr), which reduced both perchlorate and chlorate, while the genome of strain NSS included a gene for a distinct chlorate reductase (Clr) that reduced only chlorate. When both (per)chlorate and nitrate were present, (per)chlorate was preferentially utilized if the inoculum was pregrown on (per)chlorate. Historically, (per)chlorate-reducing bacteria (PRB) and chlorate-reducing bacteria (CRB) have been isolated primarily from freshwater, mesophilic environments. This study describes the isolation and characterization of two highly related marine halophiles, one a PRB and the other a CRB, and thus broadens the known phylogenetic and physiological diversity of these unusual metabolisms. Copyright © 2015, American Society for Microbiology. All Rights

European red list of habitats. Part 1: Marine habitats

NARCIS (Netherlands)

Gubbay, S.; Sanders, N.; Haynes, T.; Janssen, J.A.M.; Rodwell, J.R.; Nieto, A.; Garcia Criado, M.; Beal, S.; Borg, J.

2016-01-01

The European Red List of Habitats provides an overview of the risk
of collapse (degree of endangerment) of marine, terrestrial and
freshwater habitats in the European Union (EU28) and adjacent
regions (EU28+), based on a consistent set of categories and
criteria, and detailed data

Faunistics (marine animals)

DEFF Research Database (Denmark)

2009-01-01

These PowerPoint files are compiled from various sources: Internet, field guides, scientific monographs, textbooks, my own photos and drawings, etc. I have no copyright or permission to use most of the illustrations. The file is therefore only intended for internal use within the Marine Biology...... for identification have only been included for about a quarter of the species only, because of lack of time).     These files contain information of about 570 species of marine invertebrates found in the waters around Denmark. They should be the most common species. Which species should be selected for files like......) with the programme PowerPoint X for Mac® Service Release 1.     Comments and suggestions are welcome from students and colleagues. HD&P = Køie, Kristiansen & Weitemeyer, Havets dyr og planter. DN = Danmarks Natur, vol. 3, Havet     Tomas Cedhagen, Department of Marine Ecology, University of Aarhus, Finlandsgade 14...

Role of Diatoms in marine biofouling

Digital Repository Service at National Institute of Oceanography (India)

Anil, A; Patil, J.S..; Mitbavkar, S.; DeCosta, P.M.; DeSilva, S.; Hegde, S.; Naik, R.

. Ltd., New Delhi, pp. 293-6. de Nys, R., Leya, T., Maximilien, R., Afsar, A., Nair, P. S. R. & Steinberg, P. D. 1996. The need for standardized broad scale bioassay testing: a case study using the red alga Laurencia rigida. Biofouling 10:213-24. de...-1 Content-Type text/plain; charset=ISO-8859-1 Recent Advances on Applied Aspects of Indian Marine Algae with Reference to Global Scenario, Volume 1, A. Tewari (Ed.), 2006 Central Salt & Marine Chemicals Research Institute Role of Diatoms...

Time-Resolved Transposon Insertion Sequencing Reveals Genome-Wide Fitness Dynamics during Infection

Directory of Open Access Journals (Sweden)

Guanhua Yang

2017-10-01

Full Text Available Transposon insertion sequencing (TIS is a powerful high-throughput genetic technique that is transforming functional genomics in prokaryotes, because it enables genome-wide mapping of the determinants of fitness. However, current approaches for analyzing TIS data assume that selective pressures are constant over time and thus do not yield information regarding changes in the genetic requirements for growth in dynamic environments (e.g., during infection. Here, we describe structured analysis of TIS data collected as a time series, termed pattern analysis of conditional essentiality (PACE. From a temporal series of TIS data, PACE derives a quantitative assessment of each mutant’s fitness over the course of an experiment and identifies mutants with related fitness profiles. In so doing, PACE circumvents major limitations of existing methodologies, specifically the need for artificial effect size thresholds and enumeration of bacterial population expansion. We used PACE to analyze TIS samples of Edwardsiella piscicida (a fish pathogen collected over a 2-week infection period from a natural host (the flatfish turbot. PACE uncovered more genes that affect E. piscicida’s fitness in vivo than were detected using a cutoff at a terminal sampling point, and it identified subpopulations of mutants with distinct fitness profiles, one of which informed the design of new live vaccine candidates. Overall, PACE enables efficient mining of time series TIS data and enhances the power and sensitivity of TIS-based analyses.

Green Marine: An environmental program to establish sustainability in marine transportation.

Science.gov (United States)

Walker, Tony R

2016-04-15

European maritime companies have adopted programs to limit operational impacts on the environment. For maritime companies in North America, the Green Marine Environmental Program (GMEP) offers a framework to establish and reduce environmental footprints. Green Marine (GM) participants demonstrate annual improvements of specific environmental performance indicators (e.g., reductions in air pollution emissions) to maintain certification. Participants complete annual self-evaluations with results determining rankings for performance indicators on a 1-to-5 scale. Self-evaluations are independently verified every two years to ensure rigor and individual results are made publicly available annually to achieve transparency. GM benefits the marine industry across North America by encouraging sustainable development initiatives. GM's credibility is reflected through a diverse network of environmental groups and government agencies that endorse and help shape the program. Merits of this relatively new maritime certification (not previously described in the academic literature), are discussed. Copyright © 2016 Elsevier Ltd. All rights reserved.

A Traceless Selection: Counter-selection System That Allows Efficient Generation of Transposon and CRISPR-modified T-cell Products

Directory of Open Access Journals (Sweden)

Riccardo Mezzadra

2016-01-01

Full Text Available Recent years have seen major breakthroughs in genome-engineering systems, such as transposon-mediated gene delivery systems and CRISPR-Cas9-mediated genome-editing tools. In these systems, transient expression of auxiliary genes is responsible for permanent genomic modification. For both systems, it would be valuable to select for cells that are likely to undergo stable genome modification. Importantly, in particular for clinical applications of genome-engineered cell products, it will also be of importance to remove those cells that, due to random vector integration, display an unwanted stable expression of the auxiliary gene. Here, we develop a traceless selection system that on the one hand allows efficient enrichment of modified cells, and on the other hand can be used to select against cells that retain expression of the auxiliary gene. The value of this system to produce highly enriched-auxiliary gene-free cell products is demonstrated.

Invariability of Central Metabolic Flux Distribution in Shewanella oneidensis MR-1 Under Environmental or Genetic Perturbations

Energy Technology Data Exchange (ETDEWEB)

Tang, Yinjie; Martin, Hector Garcia; Deutschbauer, Adam; Feng, Xueyang; Huang, Rick; Llora, Xavier; Arkin, Adam; Keasling, Jay D.

2009-04-21

An environmentally important bacterium with versatile respiration, Shewanella oneidensis MR-1, displayed significantly different growth rates under three culture conditions: minimal medium (doubling time {approx} 3 hrs), salt stressed minimal medium (doubling time {approx} 6 hrs), and minimal medium with amino acid supplementation (doubling time {approx}1.5 hrs). {sup 13}C-based metabolic flux analysis indicated that fluxes of central metabolic reactions remained relatively constant under the three growth conditions, which is in stark contrast to the reported significant changes in the transcript and metabolite profiles under various growth conditions. Furthermore, ten transposon mutants of S. oneidensis MR-1 were randomly chosen from a transposon library and their flux distributions through central metabolic pathways were revealed to be identical, even though such mutational processes altered the secondary metabolism, for example, glycine and C1 (5,10-Me-THF) metabolism.

Polonium ({sup 210}Po) and lead ({sup 210}Pb) in marine organisms and their transfer in marine food chains

Energy Technology Data Exchange (ETDEWEB)

Carvalho, Fernando P., E-mail: carvalho@itn.p [Instituto Tecnologico e Nuclear, Departamento de Proteccao Radiologica e Seguranca Nuclear, E.N. 10, 2686-953 Sacavem (Portugal)

2011-05-15

The determination of {sup 210}Po and {sup 210}Pb was performed in marine organisms from the seashore to abyssal depths, encompassing a plethora of species from the microscopic plankton to the sperm whale. Concentrations of those radionuclides ranged from low values of about 5 x 10{sup -1} Bq kg{sup -1} (wet wt.) in jellyfish, to very high values of about of 3 x 10{sup 4} Bq kg{sup -1} (wet wt.) in the gut walls of sardines, with a common pattern of {sup 210}Po > {sup 210}Pb.These radionuclides are primarily absorbed from water and concentrated by phyto- and microzooplankton, and then are transferred to the next trophic level along marine food chains. Investigation in epipelagic, mesopelagic, bathypelagic and abyssobenthic organisms revealed that {sup 210}Po is transferred in the marine food webs with transfer factors ranging from 0.1 to 0.7, and numerically similar to those of the energy transfer in the marine food chains. As {sup 210}Po preferentially binds to amino acids and proteins, its transfer in food chains likely traces protein transfer and, thus, {sup 210}Po transfer factors are similar to ecotrophic coefficients. {sup 210}Pb is transferred less efficiently in marine food chains and this contributes to increased {sup 210}Po:{sup 210}Pb activity ratios in some trophic levels.

European coatings conference - Marine coatings. Proceedings

Energy Technology Data Exchange (ETDEWEB)

NONE

2008-07-01

This volume contains 13 lectures (manuscripts or powerpoint foils) with the following topics: 1. Impact of containerization on polyurethane and polyurea in marine and protective coatings (Malte Homann); 2. The application of combinatorial/high-throughput methods to the development of marine coatings (Bret Chisholm); 3. Progress and perspectives in the AMBIO (advanced nanostructured surfaces for the control of biofouling) Project (James Callow); 4. Release behaviour due to shear and pull-off of silicone coatings with a thickness gradient (James G. Kohl); 5. New liquid rheology additives for high build marine coatings (Andreas Freytag); 6. Effective corrosion protection with polyaniline, polpyrrole and polythiophene as anticorrosice additives for marine paints (Carlos Aleman); 7. Potential applications of sol gel technology for marine applications (Robert Akid); 8: Performance of biocide-free Antifouling Coatings for leisure boats (Bernd Daehne); 9. Novel biocidefree nanostructured antifouling coatings - can nano do the job? (Corne Rentrop); 10. One component high solids, VOC compliant high durability finish technology (Adrian Andrews); 11. High solid coatings - the hybrid solution (Luca Prezzi); 12. Unique organofunctional silicone resins for environmentally friendly high-performance coatings (Dieter Heldmann); 13. Silicone-alkyd paints for marine applications: from battleship-grey to green (Thomas Easton).

Marine Geophysics: a Navy Symposium

Science.gov (United States)

1987-09-01

Harrison The Source of Marine Magnetic Anomalies Christopher GA. Harrison • 52 Principles of Operation and Applications of RF-driven SQUID Magnetometers... ink recorder, and thereby obtained detailed data over the limited range in which Layer 2 appeared as a first arrival. Those researchers who waited to... Extract from MPL [Marine Physical Laboratory of the Scripps Institution of Oceanography] Quarterly Reports, 1 April to 30 June 1949, by R. W. Raitt

Marine Environmental History

DEFF Research Database (Denmark)

Poulsen, Bo

2012-01-01

human society and natural marine resources. Within this broad topic, several trends and objectives are discernable. The essay argue that the so-called material marine environmental history has its main focus on trying to reconstruct the presence, development and environmental impact of past fisheries......This essay provides an overview of recent trends in the historiography of marine environmental history, a sub-field of environmental history which has grown tremendously in scope and size over the last c. 15 years. The object of marine environmental history is the changing relationship between...... and whaling operations. This ambition often entails a reconstruction also of how marine life has changed over time. The time frame rages from Paleolithicum to the present era. The field of marine environmental history also includes a more culturally oriented environmental history, which mainly has come...

Praziquantel degradation in marine aquarium water

Directory of Open Access Journals (Sweden)

Amber Thomas

2016-04-01

Full Text Available Praziquantel (PZQ is a drug commonly utilized to treat both human schistosomiasis and some parasitic infections and infestations in animals. In the aquarium industry, PZQ can be administered in a “bath” to treat the presence of ectoparasites on both the gills and skin of fish and elasmobranchs. In order to fully treat an infestation, the bath treatment has to maintain therapeutic levels of PZQ over a period of days or weeks. It has long been assumed that, once administered, PZQ is stable in a marine environment throughout the treatment interval and must be mechanically removed, but no controlled experiments have been conducted to validate that claim. This study aimed to determine if PZQ would break down naturally within a marine aquarium below its 2 ppm therapeutic level during a typical 30-day treatment: and if so, does the presence of fish or the elimination of all living biological material impact the degradation of PZQ? Three 650 L marine aquarium systems, each containing 12 fish (French grunts: Haemulon flavolineatum, and three 650 L marine aquariums each containing no fish were treated with PZQ (2 ppm and concentrations were measured daily for 30 days. After one round of treatment, the PZQ was no longer detectable in any system after 8 (±1 days. The subsequent two PZQ treatments yielded even faster PZQ breakdown (non-detectable after 2 days and 2 ± 1 day, respectively with slight variations between systems. Linear mixed effects models of the data indicate that day and trial most impact PZQ degradation, while the presence of fish was not a factor in the best-fit models. In a completely sterilized marine system (0.5 L PZQ concentration remained unchanged over 15 days, suggesting that PZQ may be stable in a marine system during this time period. The degradation observed in non-sterile marine systems in this study may be microbial in nature. This work should be taken into consideration when providing PZQ bath treatments to marine animals

Mimicking Seawater For Culturing Marine Bacteria

DEFF Research Database (Denmark)

Rygaard, Anita Mac; Sonnenschein, Eva; Gram, Lone

2015-01-01

Only about 1% of marine bacteria have been brought into culture using traditional techniques. The purpose of this study was to investigate if mimicking the natural bacterial environment can increase culturability.We used marine substrates containing defined algal polymers or gellan gum as solidif......Only about 1% of marine bacteria have been brought into culture using traditional techniques. The purpose of this study was to investigate if mimicking the natural bacterial environment can increase culturability.We used marine substrates containing defined algal polymers or gellan gum...... as solidifying agents, and enumerated bacteria from seawater and algal exudates. We tested if culturability could be influenced by addition of quorum sensing signals (AHLs). All plates were incubated at 15°C. Bacterial counts (CFU/g) from algal exudates from brown algae were highest on media containing algal...... polymers. In general, bacteria isolated from algal exudates preferred more rich media than bacteria isolated from seawater. Overall, culturability ranged from 0.01 to 0.8% as compared to total cell count. Substitution of agar with gellan gum increased the culturability of seawater bacteria approximately...

Biological cycling of elements and stable isotopes in marine environments. Progress report, April 1, 1972-May 1, 1973

Energy Technology Data Exchange (ETDEWEB)

Kaplan, I. R.

1973-01-01

Research effort was directed toward four projects: development of a rapid technique for the measurement of total uranium content in marine sediments; understanding of the quantitative aspects involved in the anaerobic degradation of organic matter on the sea floor; evaluation of the magnitude of ionic diffusion constants in marine sediments; and development of a rapid monitoring system for dissolved sulfate in sea water and sediments to determine pollution of the ocean floor.

Proteomic characterization of the outer membrane vesicle of the halophilic marine bacterium Novosphingobium pentaromativorans US6-1.

Science.gov (United States)

Yun, Sung Ho; Lee, Sang-Yeop; Choi, Chi-Won; Lee, Hayoung; Ro, Hyun-Joo; Jun, Sangmi; Kwon, Yong Min; Kwon, Kae Kyoung; Kim, Sang-Jin; Kim, Gun-Hwa; Kim, Seung Il

2017-01-01

Novosphingobium pentaromativorans US6-1 is a Gram-negative halophilic marine bacterium able to utilize several polycyclic aromatic hydrocarbons such as phenanthrene, pyrene, and benzo[a]pyrene. In this study, using transmission electron microscopy, we confirmed that N. pentaromativorans US6-1 produces outer membrane vesicles (OMVs). N. pentaromativorans OMVs (hereafter OMV Novo ) are spherical in shape, and the average diameter of OMV Novo is 25-70 nm. Proteomic analysis revealed that outer membrane proteins and periplasmic proteins of N. pentaromativorans are the major protein components of OMV Novo . Comparative proteomic analysis with the membrane-associated protein fraction and correlation analysis demonstrated that the outer membrane proteins of OMV Novo originated from the membrane- associated protein fraction. To the best of our knowledge, this study is the first to characterize OMV purified from halophilic marine bacteria.

SERDP CU-1129 Biological Assessment for Characterizing Contamination Risk at the Genetic-, Individual-, and Population-Level

Science.gov (United States)

2004-04-14

albimanus Quetzal : a Tc1 transposon (Sum P(2) = 8.3e-07) Transposase_5, E = 5.9e-08 Topo 6-2L3 LpTc1.1 A. albimanus Quetzal : a Tc1 transposon (Sum P(2...transposon LpTc1, A. albimanus Quetzal = Anopheles albimanus Quetzal transposase (AAB02109). The clones tank and tank.1 did not have sufficient

Draft Genome Sequence of Marine Sponge Symbiont Pseudoalteromonas luteoviolacea IPB1, Isolated from Hilo, Hawaii.

Science.gov (United States)

Sakai-Kawada, Francis E; Yakym, Christopher J; Helmkampf, Martin; Hagiwara, Kehau; Ip, Courtney G; Antonio, Brandi J; Armstrong, Ellie; Ulloa, Wesley J; Awaya, Jonathan D

2016-09-22

We report here the 6.0-Mb draft genome assembly of Pseudoalteromonas luteoviolacea strain IPB1 that was isolated from the Hawaiian marine sponge Iotrochota protea Genome mining complemented with bioassay studies will elucidate secondary metabolite biosynthetic pathways and will help explain the ecological interaction between host sponge and microorganism. Copyright © 2016 Sakai-Kawada et al.

Senior High School Earth Sciences and Marine Sciences.

Science.gov (United States)

Hackenberg, Mary; And Others

This guide was developed for earth sciences and marine sciences instruction in the senior high schools of Duval County, Jacksonville, Florida. The subjects covered are: (1) Earth Science for 10th, 11th, and 12th graders; (2) Marine Biology I for 10th, 11th, and 12th graders; (3) Marine Biology II, Advanced, for 11th and 12th graders; (4) Marine…

Salmonella Persistence in Tomatoes Requires a Distinct Set of Metabolic Functions Identified by Transposon Insertion Sequencing

Science.gov (United States)

Desai, Prerak; Porwollik, Steffen; Canals, Rocio; Perez, Daniel R.; Chu, Weiping; McClelland, Michael; Teplitski, Max

2016-01-01

ABSTRACT Human enteric pathogens, such as Salmonella spp. and verotoxigenic Escherichia coli, are increasingly recognized as causes of gastroenteritis outbreaks associated with the consumption of fruits and vegetables. Persistence in plants represents an important part of the life cycle of these pathogens. The identification of the full complement of Salmonella genes involved in the colonization of the model plant (tomato) was carried out using transposon insertion sequencing analysis. With this approach, 230,000 transposon insertions were screened in tomato pericarps to identify loci with reduction in fitness, followed by validation of the screen results using competition assays of the isogenic mutants against the wild type. A comparison with studies in animals revealed a distinct plant-associated set of genes, which only partially overlaps with the genes required to elicit disease in animals. De novo biosynthesis of amino acids was critical to persistence within tomatoes, while amino acid scavenging was prevalent in animal infections. Fitness reduction of the Salmonella amino acid synthesis mutants was generally more severe in the tomato rin mutant, which hyperaccumulates certain amino acids, suggesting that these nutrients remain unavailable to Salmonella spp. within plants. Salmonella lipopolysaccharide (LPS) was required for persistence in both animals and plants, exemplifying some shared pathogenesis-related mechanisms in animal and plant hosts. Similarly to phytopathogens, Salmonella spp. required biosynthesis of amino acids, LPS, and nucleotides to colonize tomatoes. Overall, however, it appears that while Salmonella shares some strategies with phytopathogens and taps into its animal virulence-related functions, colonization of tomatoes represents a distinct strategy, highlighting this pathogen's flexible metabolism. IMPORTANCE Outbreaks of gastroenteritis caused by human pathogens have been increasingly associated with foods of plant origin, with tomatoes

Archives: Western Indian Ocean Journal of Marine Science

African Journals Online (AJOL)

Items 1 - 29 of 29 ... Archives: Western Indian Ocean Journal of Marine Science. Journal Home > Archives: Western Indian Ocean Journal of Marine Science. Log in or Register to get access to full text downloads.

33 CFR 1.01-20 - Officer in Charge, Marine Inspection.

Science.gov (United States)

2010-07-01

... that they comply with the applicable laws, rules, and regulations relating to safe construction... action in cases of misconduct, negligence, or incompetence of merchant marine officers or seamen; and the enforcement of vessel inspection, navigation, and seamen's laws in general. Specific procedures for appealing...

1,3-Oxazin-6-one Derivatives and Bohemamine-Type Pyrrolizidine Alkaloids from a Marine-Derived Streptomyces spinoverrucosus.

Science.gov (United States)

Fu, Peng; La, Scott; MacMillan, John B

2016-03-25

Two new 1,3-oxazin-6-one derivatives (1 and 2) and six new bohemamine-type pyrrolizidine alkaloids (3-8) were isolated from the marine-derived Streptomyces spinoverrucosus strain SNB-048. Their structures including the absolute configurations were fully elucidated on the basis of spectroscopic analysis, ECD spectra, quantum chemical calculations, and chemical methods. Compounds 1 and 2 possess a γ-lactam moiety and a 1,3-oxazin-6-one system.

Marine Biodiversity, Climate Change, and Governance of the Oceans

Directory of Open Access Journals (Sweden)

Robin Kundis Craig

2012-05-01

Full Text Available Governance of marine biodiversity has long suffered from lack of adequate information about the ocean’s many species and ecosystems. Nevertheless, even as we are learning much more about the ocean’s biodiversity and the impacts to it from stressors such as overfishing, habitat destruction, and marine pollution, climate change is imposing new threats and exacerbating existing threats to marine species and ecosystems. Coastal nations could vastly improve their fragmented approaches to ocean governance in order to increase the protections for marine biodiversity in the climate change era. Specifically, three key governance improvements would include: (1 incorporation of marine spatial planning as a key organizing principle of marine governance; (2 working to increase the resilience of marine ecosystems be reducing or eliminating existing stressors on those ecosystems; and (3 anticipation of climate change’s future impacts on marine biodiversity through the use of anticipatory zoning and more precautionary regulation.

Processing Marine Gravity Data Around Korea

Science.gov (United States)

Lee, Y.; Choi, K.; Kim, Y.; Ahn, Y.; Chang, M.

2008-12-01

In Korea currently 4 research ships are under operating in Korea, after the first research vessel equipped shipborne gravity meter was introduced in 1990s. These are Onnuri(launch 1991) of KORDI(Korea Ocean Research & Development Institute), Haeyang2000(launch 1996), Badaro1(launch 2002) of NORI(National Oceanographic Research Institute) and Tamhae2(launch 1997) of KIGAM(Korea Institute of Geoscience and Mineral Resources). Those of research vessel, Haeyang2000 have observed marine gravity data over 150,000 points each year from year 1996 to year 2003. Haeyang2000, about 2,500 tons, is unable to operate onshore so NORI has constructed another 600 tons research ship Badaro1 that has observed marine gravity data onshore since year 2002. Haeyang2000 finished observing marine gravity data offshore within Korean territorial waters until year 2003. Currently Badaro1 is observing marine gravity data onshore. These shipborne gravity data will be very useful and important on geodesy and geophysics research also those data can make a contribution to developing these studies. In this study NORI's shipbrne gravity data from 1996 to 2007 has been processed for fundamental data to compute Korean precise geoid. Marine gravity processing steps as followed. 1. Check the time sequence, latitude and longitude position, etc. of shipborne gravity data 2. Arrangement of the tide level below the pier and meter drift correction of each cruise. 3. Elimination of turning points. 4. The time lag correction. 5. Computation of RV's velocities, Heading angles and the Eötvös correction. 6. Kalman filtering of GPS navigation data using cross-over points. 7. Cross-over correction using least square adjustment. About 2,058,000 points have been processed with NORI's marine gravity data from 1996 to 2007 in this study. The distribution of free-air anomalies was -41.0 mgal to 136.0 mgal(mean 8.90mgal) within Korean territorial waters. The free-air anomalies processed with the marine gravity data are

Marine Corps Private Cloud Computing Environment Strategy

Science.gov (United States)

2012-05-15

leveraging economies of scale through the MCEITS PCCE, the Marine Corps will measure consumed IT resources more effectively, increase or decrease...flexible broad network access, resource pooling, elastic provisioning and measured services. By leveraging economies of scale the Marine Corps will be able...IaaS SaaS / IaaS 1 1 LCE I ACE Dets I I I I ------------------~ GIG / CJ Internet Security Boundary MCEN I DISN r :------------------ MCEN

Marine polar steroids

International Nuclear Information System (INIS)

Stonik, Valentin A

2001-01-01

Structures, taxonomic distribution and biological activities of polar steroids isolated from various marine organisms over the last 8-10 years are considered. The peculiarities of steroid biogenesis in the marine biota and their possible biological functions are discussed. Syntheses of some highly active marine polar steroids are described. The bibliography includes 254 references.

78 FR 21112 - Marine Mammals; File Nos. 16992 and 14535

Science.gov (United States)

2013-04-09

... hereby given that (1) Paul Nachtigall, Ph.D., Hawaii Institute of Marine Biology, University of Hawaii, P... (Pseudorca crassidens) maintained in captivity at the Hawaii Institute of Marine Biology in Kaneohe, HI... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration RIN 0648-XB161 Marine...

An assessment of natural product discovery from marine (sensu strictu) and marine-derived fungi

Science.gov (United States)

Overy, David P.; Bayman, Paul; Kerr, Russell G.; Bills, Gerald F.

2014-01-01

The natural products community has been investigating secondary metabolites from marine fungi for several decades, but when one attempts to search for validated reports of new natural products from marine fungi, one encounters a literature saturated with reports from ‘marine-derived’ fungi. Of the 1000+ metabolites that have been characterized to date, only approximately 80 of these have been isolated from species from exclusively marine lineages. These metabolites are summarized here along with the lifestyle and habitats of their producing organisms. Furthermore, we address some of the reasons for the apparent disconnect between the stated objectives of discovering new chemistry from marine organisms and the apparent neglect of the truly exceptional obligate marine fungi. We also offer suggestions on how to reinvigorate enthusiasm for marine natural products discovery from fungi from exclusive marine lineages and highlight the need for critically assessing the role of apparently terrestrial fungi in the marine environment. PMID:25379338

A role for the budding yeast separase, Esp1, in Ty1 element retrotransposition.

Directory of Open Access Journals (Sweden)

Krystina L Ho

2015-03-01

Full Text Available Separase/Esp1 is a protease required at the onset of anaphase to cleave cohesin and thereby enable sister chromatid separation. Esp1 also promotes release of the Cdc14 phosphatase from the nucleolus to enable mitotic exit. To uncover other potential roles for separase, we performed two complementary genome-wide genetic interaction screens with a strain carrying the budding yeast esp1-1 separase mutation. We identified 161 genes that when mutated aggravate esp1-1 growth and 44 genes that upon increased dosage are detrimental to esp1-1 viability. In addition to the expected cell cycle and sister chromatid segregation genes that were identified, 24% of the genes identified in the esp1-1 genetic screens have a role in Ty1 element retrotransposition. Retrotransposons, like retroviruses, replicate through reverse transcription of an mRNA intermediate and the resultant cDNA product is integrated into the genome by a conserved transposon or retrovirus encoded integrase protein. We purified Esp1 from yeast and identified an interaction between Esp1 and Ty1 integrase using mass spectrometry that was subsequently confirmed by co-immunoprecipitation analysis. Ty1 transposon mobility and insertion upstream of the SUF16 tRNA gene are both reduced in an esp1-1 strain but increased in cohesin mutant strains. Securin/Pds1, which is required for efficient localization of Esp1 to the nucleus, is also required for efficient Ty1 transposition. We propose that Esp1 serves two roles to mediate Ty1 transposition - one to remove cohesin and the second to target Ty1-IN to chromatin.

Long interspersed element-1 (LINE-1): passenger or driver in human neoplasms?

Science.gov (United States)

Rodić, Nemanja; Burns, Kathleen H

2013-03-01

LINE-1 (L1) retrotransposons make up a significant portion of human genomes, with an estimated 500,000 copies per genome. Like other retrotransposons, L1 retrotransposons propagate through RNA sequences that are reverse transcribed into DNA sequences, which are integrated into new genomic loci. L1 somatic insertions have the potential to disrupt the transcriptome by inserting into or nearby genes. By mutating genes and playing a role in epigenetic dysregulation, L1 transposons may contribute to tumorigenesis. Studies of the "mobilome" have lagged behind other tumor characterizations at the sequence, transcript, and epigenetic levels. Here, we consider evidence that L1 retrotransposons may sometimes drive human tumorigenesis.

Marine Mammals: Hearing and Echolocation at Coconut Island

Science.gov (United States)

2012-09-30

1 DISTRIBUTION STATEMENT A. Approved for public release; distribution is unlimited. Marine Mammals: Hearing and Echolocation at Coconut ...REPORT DATE 2012 2. REPORT TYPE N/A 3. DATES COVERED - 4. TITLE AND SUBTITLE Marine Mammals: Hearing and Echolocation at Coconut Island

Public perspective towards marine litter in West Aceh City

Science.gov (United States)

Kusumawati, I.; Setyowati, M.; Riana, E.; Prartono, T.

2018-03-01

Marine litter or marine debris is a man-made solid material discarded, abandoned or lost in coastline or into the sea. To reduce the amount of marine litter in the ocean, raising public awareness is an important way. One of the contributing factors on marine litter is the lack of understanding within the community, but to identify how people notice the problem is required adequate research literature. The purpose of this study is to examine the awareness of West Aceh community on marine litter along western coastal area. The research objectives; 1) to evaluate societal perception towards marine litter; 2) to examine the urgent indicator of public awareness in West Aceh City. This study will employ a survey approach by distributing questionnaires to 383 respondents. It was found that respondents show low awareness on marine litter according to statistical data, but there are some rooms to manage in order to raise the level of public awareness. It concludes that sense of responsibility could be enhanced by involving public in any activities for preventing and eradicating marine litter. Education aspect is also important to increase public understanding about the threats of marine debris on environment, human health and economic income.

Marine disease impacts, diagnosis, forecasting, management and policy

Science.gov (United States)

Lafferty, Kevin D.; Hofmann, Eileen E.

2016-01-01

As Australians were spending millions of dollars in 2014 to remove the coral-eating crown of thorns sea star from the Great Barrier Reef, sea stars started washing up dead for free along North America's Pacific Coast. Because North American sea stars are important and iconic predators in marine communities, locals and marine scientists alike were alarmed by what proved to be the world's most widespread marine mass mortality in geographical extent and species affected, especially given its mysterious cause. Investigative research using modern diagnostic techniques implicated a never-before-seen virus [1]. The virus inspired international attention to marine diseases, including this theme issue.

Autoantigen La promotes efficient RNAi, antiviral response, and transposon silencing by facilitating multiple-turnover RISC catalysis.

Science.gov (United States)

Liu, Ying; Tan, Huiling; Tian, Hui; Liang, Chunyang; Chen, She; Liu, Qinghua

2011-11-04

The effector of RNA interference (RNAi) is the RNA-induced silencing complex (RISC). C3PO promotes the activation of RISC by degrading the Argonaute2 (Ago2)-nicked passenger strand of duplex siRNA. Active RISC is a multiple-turnover enzyme that uses the guide strand of siRNA to direct the Ago2-mediated sequence-specific cleavage of complementary mRNA. How this effector step of RNAi is regulated is currently unknown. Here, we used the human Ago2 minimal RISC system to purify Sjögren's syndrome antigen B (SSB)/autoantigen La as an activator of the RISC-mediated mRNA cleavage activity. Our reconstitution studies showed that La could promote multiple-turnover RISC catalysis by facilitating the release of cleaved mRNA from RISC. Moreover, we demonstrated that La was required for efficient RNAi, antiviral defense, and transposon silencing in vivo. Taken together, the findings of C3PO and La reveal a general concept that regulatory factors are required to remove Ago2-cleaved products to assemble or restore active RISC. Copyright © 2011 Elsevier Inc. All rights reserved.

ELIXIR pilot action: Marine metagenomics – towards a domain specific set of sustainable services [version 1; referees: 1 approved, 2 approved with reservations

Directory of Open Access Journals (Sweden)

Espen Mikal Robertsen

2017-01-01

Full Text Available Metagenomics, the study of genetic material recovered directly from environmental samples, has the potential to provide insight into the structure and function of heterogeneous microbial communities.  There has been an increased use of metagenomics to discover and understand the diverse biosynthetic capacities of marine microbes, thereby allowing them to be exploited for industrial, food, and health care products. This ELIXIR pilot action was motivated by the need to establish dedicated data resources and harmonized metagenomics pipelines for the marine domain, in order to enhance the exploration and exploitation of marine genetic resources. In this paper, we summarize some of the results from the ELIXIR pilot action “Marine metagenomics – towards user centric services”.

Recovery trends in marine mammal populations.

Directory of Open Access Journals (Sweden)

Anna M Magera

Full Text Available Marine mammals have greatly benefitted from a shift from resource exploitation towards conservation. Often lauded as symbols of conservation success, some marine mammal populations have shown remarkable recoveries after severe depletions. Others have remained at low abundance levels, continued to decline, or become extinct or extirpated. Here we provide a quantitative assessment of (1 publicly available population-level abundance data for marine mammals worldwide, (2 abundance trends and recovery status, and (3 historic population decline and recent recovery. We compiled 182 population abundance time series for 47 species and identified major data gaps. In order to compare across the largest possible set of time series with varying data quality, quantity and frequency, we considered an increase in population abundance as evidence of recovery. Using robust log-linear regression over three generations, we were able to classify abundance trends for 92 spatially non-overlapping populations as Significantly Increasing (42%, Significantly Decreasing (10%, Non-Significant Change (28% and Unknown (20%. Our results were comparable to IUCN classifications for equivalent species. Among different groupings, pinnipeds and other marine mammals (sirenians, polar bears and otters showed the highest proportion of recovering populations, likely benefiting from relatively fast life histories and nearshore habitats that provided visibility and protective management measures. Recovery was less frequent among cetaceans, but more common in coastal than offshore populations. For marine mammals with available historical abundance estimates (n = 47, larger historical population declines were associated with low or variable recent recoveries so far. Overall, our results show that many formerly depleted marine mammal populations are recovering. However, data-deficient populations and those with decreasing and non-significant trends require attention. In particular

Carotenoids in Marine Animals

Science.gov (United States)

Maoka, Takashi

2011-01-01

Marine animals contain various carotenoids that show structural diversity. These marine animals accumulate carotenoids from foods such as algae and other animals and modify them through metabolic reactions. Many of the carotenoids present in marine animals are metabolites of β-carotene, fucoxanthin, peridinin, diatoxanthin, alloxanthin, and astaxanthin, etc. Carotenoids found in these animals provide the food chain as well as metabolic pathways. In the present review, I will describe marine animal carotenoids from natural product chemistry, metabolism, food chain, and chemosystematic viewpoints, and also describe new structural carotenoids isolated from marine animals over the last decade. PMID:21566799

Marine Biomedicine

Science.gov (United States)

Bang, Frederik B.

1977-01-01

Describes early scientific research involving marine invertebrate pathologic processes that may have led to new insights into human disease. Discussed are inquiries of Metchnikoff, Loeb, and Cantacuzene (immunolgic responses in sea stars, horseshoe crabs, and marine worms, respectively). Describes current research stemming from these early…

Marine Biology

Science.gov (United States)

Dewees, Christopher M.; Hooper, Jon K.

1976-01-01

A variety of informational material for a course in marine biology or oceanology at the secondary level is presented. Among the topics discussed are: food webs and pyramids, planktonic blooms, marine life, plankton nets, food chains, phytoplankton, zooplankton, larval plankton and filter feeders. (BT)

Active Marine Station Metadata

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Active Marine Station Metadata is a daily metadata report for active marine bouy and C-MAN (Coastal Marine Automated Network) platforms from the National Data...

Characterizing Marine Soundscapes.

Science.gov (United States)

Erbe, Christine; McCauley, Robert; Gavrilov, Alexander

2016-01-01

The study of marine soundscapes is becoming widespread and the amount of data collected is increasing rapidly. Data owners (typically academia, industry, government, and defense) are negotiating data sharing and generating potential for data syntheses, comparative studies, analyses of trends, and large-scale and long-term acoustic ecology research. A problem is the lack of standards and commonly agreed protocols for the recording of marine soundscapes, data analysis, and reporting that make a synthesis and comparison of results difficult. We provide a brief overview of the components in a marine soundscape, the hard- and software tools for recording and analyzing marine soundscapes, and common reporting formats.

African Journal of Marine Science - Vol 36, No 1 (2014)

African Journals Online (AJOL)

Optimal BRUVs (baited remote underwater video system) survey design for reef fish monitoring in the Stilbaai Marine Protected Area · EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. L De Vos, A Götz, H Winker, CG Attwood. http://dx.doi.org/10.2989/1814232X.2013.873739 ...

Carotenoids in Marine Animals

OpenAIRE

Maoka, Takashi

2011-01-01

Marine animals contain various carotenoids that show structural diversity. These marine animals accumulate carotenoids from foods such as algae and other animals and modify them through metabolic reactions. Many of the carotenoids present in marine animals are metabolites of β-carotene, fucoxanthin, peridinin, diatoxanthin, alloxanthin, and astaxanthin, etc. Carotenoids found in these animals provide the food chain as well as metabolic pathways. In the present review, I will describe marine a...

A new physically-based quantification of marine isoprene and primary organic aerosol emissions

Directory of Open Access Journals (Sweden)

N. Meskhidze

2009-07-01

Full Text Available The global marine sources of organic carbon (OC are estimated here using a physically-based parameterization for the emission of marine isoprene and primary organic matter. The marine isoprene emission model incorporates new physical parameters such as light sensitivity of phytoplankton isoprene production and dynamic euphotic depth to simulate hourly marine isoprene emissions totaling 0.92 Tg C yr⁻¹. Sensitivity studies using different schemes for the euphotic zone depth and ocean phytoplankton speciation produce the upper and the lower range of marine-isoprene emissions of 0.31 to 1.09 Tg C yr⁻¹, respectively. Established relationships between sea spray fractionation of water-insoluble organic carbon (WIOC and chlorophyll-a concentration are used to estimate the total primary sources of marine sub- and super-micron OC of 2.9 and 19.4 Tg C yr⁻¹, respectively. The consistent spatial and temporal resolution of the two emission types allow us, for the first time, to explore the relative contributions of sub- and super-micron organic matter and marine isoprene-derived secondary organic aerosol (SOA to the total OC fraction of marine aerosol. Using a fixed 3% mass yield for the conversion of isoprene to SOA, our emission simulations show minor (<0.2% contribution of marine isoprene to the total marine source of OC on a global scale. However, our model calculations also indicate that over the tropical oceanic regions (30° S to 30° N, marine isoprene SOA may contribute over 30% of the total monthly-averaged sub-micron OC fraction of marine aerosol. The estimated contribution of marine isoprene SOA to hourly-averaged sub-micron marine OC emission is even higher, approaching 50% over the vast regions of the oceans during the midday hours when isoprene emissions are highest. As it is widely believed that sub-micron OC has the potential to influence the cloud droplet activation of marine aerosols, our

Marine Science

African Journals Online (AJOL)

Aims and scope: The Western Indian Ocean Journal of Marine Science provides an avenue for the wide dissem- ination of high ... or by any means without permission in writing from the copyright holder. ..... Journal of Chemical Engineering Research and Design 82 ... Indian Ocean Marine Science Association Technical.

Nitrogen and phosphorus treatment of marine wastewater by a laboratory-scale sequencing batch reactor with eco-friendly marine high-efficiency sediment.

Science.gov (United States)

Cho, Seonghyeon; Kim, Jinsoo; Kim, Sungchul; Lee, Sang-Seob

2017-06-22

We screened and identified a NH 3 -N-removing bacterial strain, Bacillus sp. KGN1, and a [Formula: see text] removing strain, Vibrio sp. KGP1, from 960 indigenous marine isolates from seawater and marine sediment from Tongyeong, South Korea. We developed eco-friendly high-efficiency marine sludge (eco-HEMS), and inoculated these marine bacterial strains into the marine sediment. A laboratory-scale sequencing batch reactor (SBR) system using the eco-HEMS for marine wastewater from land-based fish farms improved the treatment performance as indicated by 88.2% removal efficiency (RE) of total nitrogen (initial: 5.6 mg/L) and 90.6% RE of total phosphorus (initial: 1.2 mg/L) under the optimal operation conditions (food and microorganism (F/M) ratio, 0.35 g SCOD Cr /g mixed liquor volatile suspended solids (MLVSS)·d; dissolved oxygen (DO) 1.0 ± 0.2 mg/L; hydraulic retention time (HRT), 6.6 h; solids retention time (SRT), 12 d). The following kinetic parameters were obtained: cell yield (Y), 0.29 g MLVSS/g SCOD Cr ; specific growth rate (µ), 0.06 d -1 ; specific nitrification rate (SNR), 0.49 mg NH 3 -N/g MLVSS·h; specific denitrification rate (SDNR), 0.005 mg [Formula: see text]/g MLVSS·h; specific phosphorus uptake rate (SPUR), 0.12 mg [Formula: see text]/g MLVSS·h. The nitrogen- and phosphorus-removing bacterial strains comprised 18.4% of distribution rate in the microbial community of eco-HEMS under the optimal operation conditions. Therefore, eco-HEMS effectively removed nitrogen and phosphorus from highly saline marine wastewater from land-based fish farms with improving SNR, SDNR, and SPUR values in more diverse microbial communities. DO: dissolved oxygen; Eco-HEMS: eco-friendly high efficiency marine sludge; F/M: food and microorganism ratio; HRT: hydraulic retention time; ML(V)SS: mixed liquor (volatile) suspended solids; NCBI: National Center for Biotechnology Information; ND: not determined; qPCR: quantitative real-time polymerase

Russian spent marine fuel as a global security risk

International Nuclear Information System (INIS)

Gussgard, K.; Reistad, O.

2001-01-01

Russian marine fuel is a trans-national security concern. This paper focuses on specific technical properties of Russian marine nuclear fuel especially relevant for evaluating different aspects on nuclear proliferation, in addition to risks associated with regional environmental degradation and illegal diversion of radiological substances. Russian fresh fuel for marine reactors has been involved in several significant cases of illicit trafficking of special nuclear materials. The amount and quality of nuclear materials in Russian spent marine fuel give also reason for concern. Not less than 200 marine reactor cores are ready for having their spent fuel unloaded and preliminary stored on shore in the Far East and North West of Russia, and large amounts of spent naval fuel have been stored at Russian military bases for decades. In order to assess the security risks associated with Russian spent marine fuel, this paper discusses the material attractiveness of spent fuel from all types of Russian marine reactors. The calculations are based on a model of a light water moderated Russian icebreaker reactor. The computer tool HELIOS, used for modelling the reactor and the reactor operations, has been extensively qualified by comparisons with experimental data and international benchmark problems for reactor physics codes as well as through feedback from applications. Some of these benchmarks and studies include fuel enrichments up to 90% in Russian marine reactors. Several fuel data cases are discussed in the paper, focusing especially on: 1) early fuel designs with low initial enrichment; 2) more modern fuel designs used in third and fourth generation of Russian submarines probably with intermediate enriched fuel; and 3) marine fuel with initial enrichment levels close to weapons-grade material. In each case the fuel has been burned until k eff has reached below 1. Case 1) has been evaluated, the calculations made as basis for this paper have concentrated on fuel with

Nitrous oxide production associated with coastal marine invertebrates

DEFF Research Database (Denmark)

Heisterkamp, Ines Maria; Schramm, Andreas; de Beer, Dirk

2010-01-01

Several freshwater and terrestrial invertebrate species emit the greenhouse gas nitrous oxide (N2O). The N2O production associated with these animals was ascribed to incomplete denitrification by ingested sediment or soil bacteria. The present study shows that many marine invertebrates also emit N2......O at substantial rates. A total of 19 invertebrate species collected in the German Wadden Sea and in Aarhus Bay, Denmark, and 1 aquacultured shrimp species were tested for N2O emission. Potential N2O emission rates ranged from 0 to 1.354 nmol ind.–1 h–1, with an average rate of 0.320 nmol ind.–1 h–1...... with an experimentally cleaned shell. Thus, the N2O production associated with marine invertebrates is apparently not due to gut denitrification in every species, but may also result from microbial activity on the external surfaces of animals. The high abundance and potential N2O emission rates of many marine...

Marine animal stings or bites

Science.gov (United States)

Stings - marine animals; Bites - marine animals ... Things you can do to prevent a marine animal sting or bite include: Swim near a lifeguard. Observe posted signs that may warn of danger from jellyfish or other hazardous marine life. ...

Implementing and Innovating Marine Monitoring Approaches for Assessing Marine Environmental Status

KAUST Repository

Danovaro, Roberto; Carugati, Laura; Berzano, Marco; Cahill, Abigail E.; Carvalho, Susana; Chenuil, Anne; Corinaldesi, Cinzia; Cristina, Sonia; David, Romain; Dell'Anno, Antonio; Dzhembekova, Nina; Garcé s, Esther; Gasol, Joseph M.; Goela, Priscila; Fé ral, Jean-Pierre; Ferrera, Isabel; Forster, Rodney M.; Kurekin, Andrey A.; Rastelli, Eugenio; Marinova, Veselka; Miller, Peter I.; Moncheva, Snejana; Newton, Alice; Pearman, John K.; Pitois, Sophie G.; Reñ é , Albert; Rodrí guez-Ezpeleta, Naiara; Saggiomo, Vincenzo; Simis, Stefan G. H.; Stefanova, Kremena; Wilson, Christian; Lo Martire, Marco; Greco, Silvestro; Cochrane, Sabine K. J.; Mangoni, Olga; Borja, Angel

2016-01-01

Marine environmental monitoring has tended to focus on site-specific methods of investigation. These traditional methods have low spatial and temporal resolution and are relatively labor intensive per unit area/time that they cover. To implement the Marine Strategy Framework Directive (MSFD), European Member States are required to improve marine monitoring and design monitoring networks. This can be achieved by developing and testing innovative and cost-effective monitoring systems, as well as indicators of environmental status. Here, we present several recently developed methodologies and technologies to improve marine biodiversity indicators and monitoring methods. The innovative tools are discussed concerning the technologies presently utilized as well as the advantages and disadvantages of their use in routine monitoring. In particular, the present analysis focuses on: (i) molecular approaches, including microarray, Real Time quantitative PCR (qPCR), and metagenetic (metabarcoding) tools; (ii) optical (remote) sensing and acoustic methods; and (iii) in situ monitoring instruments. We also discuss their applications in marine monitoring within the MSFD through the analysis of case studies in order to evaluate their potential utilization in future routine marine monitoring. We show that these recently-developed technologies can present clear advantages in accuracy, efficiency and cost.

Implementing and Innovating Marine Monitoring Approaches for Assessing Marine Environmental Status

KAUST Repository

Danovaro, Roberto

2016-11-23

Marine environmental monitoring has tended to focus on site-specific methods of investigation. These traditional methods have low spatial and temporal resolution and are relatively labor intensive per unit area/time that they cover. To implement the Marine Strategy Framework Directive (MSFD), European Member States are required to improve marine monitoring and design monitoring networks. This can be achieved by developing and testing innovative and cost-effective monitoring systems, as well as indicators of environmental status. Here, we present several recently developed methodologies and technologies to improve marine biodiversity indicators and monitoring methods. The innovative tools are discussed concerning the technologies presently utilized as well as the advantages and disadvantages of their use in routine monitoring. In particular, the present analysis focuses on: (i) molecular approaches, including microarray, Real Time quantitative PCR (qPCR), and metagenetic (metabarcoding) tools; (ii) optical (remote) sensing and acoustic methods; and (iii) in situ monitoring instruments. We also discuss their applications in marine monitoring within the MSFD through the analysis of case studies in order to evaluate their potential utilization in future routine marine monitoring. We show that these recently-developed technologies can present clear advantages in accuracy, efficiency and cost.

Identification of a Marine Bacillus Strain C5 and Parathion-Methyl Degradation Characteristics of the Extracellular Esterase B1

Directory of Open Access Journals (Sweden)

Jianhua Hao

2014-01-01

Full Text Available A bacterial strain C5 that can produce new type of marine esterase was isolated and screened from marine sludge. According to 16S rRNA sequence analysis and physiological and biochemical experiments, the strain was identified as Bacillus subtilis. A single isozyme with a molecular weight of 86 kDa was observed by SDS-PAGE and native-PAGE. On this basis, the mechanism of esterase B1 secreted by strain C5 degrading parathion-methyl was explored, and the effects of temperature and pH on the degradation rate were investigated. From the results, p-nitrophenol was one of the degradation products of B1 degrading parathion-methyl, and the best degradation effect could be achieved at the temperature of 40°C and the neutral pH value.

Experimental design for three interrelated marine ice sheet and ocean model intercomparison projects: MISMIP v. 3 (MISMIP +), ISOMIP v. 2 (ISOMIP +) and MISOMIP v. 1 (MISOMIP1)

Energy Technology Data Exchange (ETDEWEB)

Asay-Davis, Xylar S. [Potsdam Inst. for Climate Impact Research (Germany). Earth System Analysis; Cornford, Stephen L. [Univ. of Bristol (United Kingdom). Centre for Polar Observation and Modelling; Durand, Gaël [Centre National de la Recherche Scientifique (CNRS), Grenoble (France); Univ. of Grenoble (France); Galton-Fenzi, Benjamin K. [Australian Antarctic Division and Antarctic Climate and Ecosystems Cooperative Research Centre, Tasmania (Australia); Gladstone, Rupert M. [Antarctic Climate and Ecosystems Cooperative Research Centre, Tasmania (Australia); ETH Zurich (Switzerland). Research Center for Hydraulic Engineering; Gudmundsson, G. Hilmar [British Antarctic Survey, Cambridge (United Kingdom); Hattermann, Tore [Akvaplan-niva, Tromso (Norway); Helmholtz Centre for Polar and Marine Research, Bremerhaven (Germany). Alfred Wegener Inst.; Holland, David M. [New York Univ. (NYU), NY (United States)' . Courant Inst. of Mathematical Sciences; Holland, Denise [New York Univ. (NYU), Abu Dahabi (United Arab Emirates); Holland, Paul R. [British Antarctic Survey, Cambridge (United Kingdom); Martin, Daniel F. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Mathiot, Pierre [British Antarctic Survey, Cambridge (United Kingdom); Met Office, Exeter (United Kingdom); Pattyn, Frank [Univ. of Libre, Brussels (Belgium). Lab. of Glaciology; Seroussi, Hélène [California Inst. of Technology (CalTech), Pasadena, CA (United States). Jet Propulsion Lab.

2016-01-01

Coupled ice sheet-ocean models capable of simulating moving grounding lines are just becoming available. Such models have a broad range of potential applications in studying the dynamics of marine ice sheets and tidewater glaciers, from process studies to future projections of ice mass loss and sea level rise. The Marine Ice Sheet-Ocean Model Intercomparison Project (MISOMIP) is a community effort aimed at designing and coordinating a series of model intercomparison projects (MIPs) for model evaluation in idealized setups, model verification based on observations, and future projections for key regions of the West Antarctic Ice Sheet (WAIS).

Here we describe computational experiments constituting three interrelated MIPs for marine ice sheet models and regional ocean circulation models incorporating ice shelf cavities. These consist of ice sheet experiments under the Marine Ice Sheet MIP third phase (MISMIP+), ocean experiments under the Ice Shelf-Ocean MIP second phase (ISOMIP+) and coupled ice sheet-ocean experiments under the MISOMIP first phase (MISOMIP1). All three MIPs use a shared domain with idealized bedrock topography and forcing, allowing the coupled simulations (MISOMIP1) to be compared directly to the individual component simulations (MISMIP+ and ISOMIP+). The experiments, which have qualitative similarities to Pine Island Glacier Ice Shelf and the adjacent region of the Amundsen Sea, are designed to explore the effects of changes in ocean conditions, specifically the temperature at depth, on basal melting and ice dynamics. In future work, differences between model results will form the basis for the evaluation of the participating models.

Marine Inspired 2-(5-Halo-1H-indol-3-yl)-N,N-dimethylethanamines as Modulators of Serotonin Receptors: An Example Illustrating the Power of Bromine as Part of the Uniquely Marine Chemical Space.

Science.gov (United States)

Ibrahim, Mohamed A; El-Alfy, Abir T; Ezel, Kelly; Radwan, Mohamed O; Shilabin, Abbas G; Kochanowska-Karamyan, Anna J; Abd-Alla, Howaida I; Otsuka, Masami; Hamann, Mark T

2017-08-09

In previous studies, we have isolated several marine indole alkaloids and evaluated them in the forced swim test (FST) and locomotor activity test, revealing their potential as antidepressant and sedative drug leads. Amongst the reported metabolites to display such activities was 5-bromo- N , N -dimethyltryptamine. Owing to the importance of the judicious introduction of halogens into drug candidates, we synthesized two series built on a 2-(1 H -indol-3-yl)- N , N -dimethylethanamine scaffold with different halogen substitutions. The synthesized compounds were evaluated for their in vitro and in vivo antidepressant and sedative activities using the mouse forced swim and locomotor activity tests. Receptor binding studies of these compounds to serotonin (5-HT) receptors were conducted. Amongst the prepared compounds, 2-(1 H -indol-3-yl)- N , N -dimethyl-2-oxoacetamide ( 1a ), 2-(5-bromo-1 H -indol-3-yl)- N , N -dimethyl-2-oxoacetamide ( 1d ), 2-(1 H -indol-3-yl)- N , N -dimethylethanamine ( 2a ), 2-(5-chloro-1 H -indol-3-yl)- N , N -dimethylethanamine ( 2c ), 2-(5-bromo-1 H -indol-3-yl)- N , N -dimethylethanamine ( 2d ), and 2-(5-iodo-1 H -indol-3-yl)- N , N -dimethylethanamine ( 2e ) have been shown to possess significant antidepressant-like action, while compounds 2c , 2d , and 2e exhibited potent sedative activity. Compounds 2a , 2c , 2d , and 2e showed nanomolar affinities to serotonin receptors 5-HT 1A and 5-HT₇. The in vitro data indicates that the antidepressant action exerted by these compounds in vivo is mediated, at least in part, via interaction with serotonin receptors. The data presented here shows the valuable role that bromine plays in providing novel chemical space and electrostatic interactions. Bromine is ubiquitous in the marine environment and a common element of marine natural products.

Seashore marine table quiz

OpenAIRE

Institute, Marine

2013-01-01

Develop an increasing awareness of plants and animals that live in local marine environments including the seashore, seas and oceans of Ireland. After learning all about the seashore and other marine related lessons, this quiz can be used to evaluate the student’s knowledge of the marine related living things and natural environments. The table quiz can be used as a guide, highlighting facts about the marine environment and some of the animals that live there.

Multiple roles of Pseudomonas aeruginosa TBCF10839 PilY1 in motility, transport and infection

DEFF Research Database (Denmark)

Bohn, Yu-Sing Tammy; Brandes, Gudrun; Rakhimova, Elza

2009-01-01

Polymorphonuclear neutrophils are the most important mammalian host defence cells against infections with Pseudomonas aeruginosa. Screening of a signature tagged mutagenesis library of the non-piliated P. aeruginosa strain TBCF10839 uncovered that transposon inactivation of its pilY1 gene rendere...

Nitrogen regulates chitinase gene expression in a marine bacterium

DEFF Research Database (Denmark)

Delpin, Marina; Goodman, A.E.

2009-01-01

Ammonium concentration and nitrogen source regulate promoter activity and use for the transcription of chiA, the major chitinase gene of Pseudoalteromonas sp. S91 and S91CX, an S91 transposon lacZ fusion mutant. The activity of chiA was quantified by beta-galactosidase assay of S91CX cultures con...... GlcNAc, transcription initiated from two putative sigma(54)-dependent promoters and (3) glt, transcription initiated from all three putative promoters. The ISME Journal (2009) 3, 1064-1069; doi:10.1038/ismej.2009.49; published online 14 May 2009...

Radioactive contamination of the Guatemalan marine environment

Energy Technology Data Exchange (ETDEWEB)

Perez-Sabino, J F; Oliva de Sandoval, B E; Orozco-Chilel, R M; Aguilar-Sandoval, E [Universidad de San Carlos de Guatemala, Facultad de Ciencias Quimicas y Farmacia, Unidad de Analisis Instrumental, Guatemala C.A. (Guatemala)

1999-07-01

As part of the IAEA TC project GUA/2/005 `Radioactivity and Contamination of the Marine Environment in Guatemala`, concentrations of artificial and natural radionuclides have been determined in marine water and sediments, giving important information to establish the base line of the natural radioactivity and the radioactive contamination in this area that not have been studying 4 refs, 1 fig., 4 tabs

Safety evaluation on MOX new fuel at marine transport

International Nuclear Information System (INIS)

Tsumune, Daisuke; Ito, Chihiro; Saegusa, Toshiari; Maruyama, Koki

2000-01-01

In the Central Research Institute of Electric Power Industry, in order to confirm effects of MOX new fuel on the public are as small as possible even when its marine transport goes down, some exposed radiation dose has previously conducted on imaginary shipwreck of marine transport on used nuclear fuel, plutonium dioxide, and high level return glass solid. Under a base of such informations, some investigations on safety on marine transport of the MOX new fuel was conducted. On September, 1999, five transport vessels of the MOX new fuel was at first transported on marine. The value of five times of estimated exposed radiation dose (max. 8.1 x 10 -8 mSv/y) corresponds to an evaluation result assumed by shipwreck in marine transport this time. As a result, it was found that the exposed radiation dose estimated on this case would be sufficiently less than an effective dose equivalent limit (1 mSv/y) of public exposure according to the recommendation of ICRP in both coastal and oceanic areas. (G.K.)

Compressibility characteristics of Sabak Bernam Marine Clay

Science.gov (United States)

Lat, D. C.; Ali, N.; Jais, I. B. M.; Baharom, B.; Yunus, N. Z. M.; Salleh, S. M.; Azmi, N. A. C.

2018-04-01

This study is carried out to determine the geotechnical properties and compressibility characteristics of marine clay collected at Sabak Bernam. The compressibility characteristics of this soil are determined from 1-D consolidation test and verified by existing correlations by other researchers. No literature has been found on the compressibility characteristics of Sabak Bernam Marine Clay. It is important to carry out this study since this type of marine clay covers large coastal area of west coast Malaysia. This type of marine clay was found on the main road connecting Klang to Perak and the road keeps experiencing undulation and uneven settlement which jeopardise the safety of the road users. The soil is indicated in the Generalised Soil Map of Peninsular Malaysia as a CLAY with alluvial soil on recent marine and riverine alluvium. Based on the British Standard Soil Classification and Plasticity Chart, the soil is classified as a CLAY with very high plasticity (CV). Results from laboratory test on physical properties and compressibility parameters show that Sabak Bernam Marine Clay (SBMC) is highly compressible, has low permeability and poor drainage characteristics. The compressibility parameters obtained for SBMC is in a good agreement with other researchers in the same field.

Developing a Capacity Assessment Framework for Marine Logistics Groups

Science.gov (United States)

2017-02-20

Corps, 2015; and 1st Marine Logistics Group, 2015. RAND RR1572-2.3 1st FSSG Headquarters Regiment Security Company CLR-1 CLR-15 7th ESB 1st Medical... Companies . Supply and maintenance battalions form the bulk of CLR-15. The Combat Logistics Companies are small, task-organized companies that support Marine...under- standing logistics capacity as it pertains to informing decisions to improve efficiencies and reduce costs . Most often these assessments of

U.S. Marine Corps Enlisted Retention: An Analysis of Stakeholder Incentives for the Retention of Tier 1 First-Term Marines

Science.gov (United States)

2014-03-01

ERTF Enlisted Retention Task Force FFM fast filling military occupational specialty FTAP first-term alignment plan FY fiscal year GT general...Planning military ( FFM ) occupational specialties that are processed through boards to ensure the retention of highly qualified Marines. Fast-filling

Marine Jurisdiction Boundaries

Data.gov (United States)

Department of Homeland Security — The NOAA Coastal Services Center's Marine Jurisdiction dataset was created to assist in marine spatial planning and offshore alternative energy sitting. This is a...

75 FR 68605 - Marine Mammals

Science.gov (United States)

2010-11-08

...-XX23 Marine Mammals AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric... permit to conduct research on marine mammals. ADDRESSES: The permit and related documents are available... applicant. The requested permit has been issued under the authority of the Marine Mammal Protection Act of...

77 FR 2512 - Marine Mammals

Science.gov (United States)

2012-01-18

...-XA905 Marine Mammals AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric... Dorian Houser, Ph.D., National Marine Mammal Foundation, 2240 Shelter Island Drive, 200, San Diego, CA... subject permit is requested under the authority of the Marine Mammal Protection Act of 1972, as amended...

Long-Term PEDF Release in Rat Iris and Retinal Epithelial Cells after Sleeping Beauty Transposon-Mediated Gene Delivery

Directory of Open Access Journals (Sweden)

Laura Garcia-Garcia

2017-12-01

Full Text Available Pigment epithelium derived factor (PEDF is a potent antiangiogenic, neurotrophic, and neuroprotective molecule that is the endogenous inhibitor of vascular endothelial growth factor (VEGF in the retina. An ex vivo gene therapy approach based on transgenic overexpression of PEDF in the eye is assumed to rebalance the angiogenic-antiangiogenic milieu of the retina, resulting in growth regression of choroidal blood vessels, the hallmark of neovascular age-related macular degeneration. Here, we show that rat pigment epithelial cells can be efficiently transfected with the PEDF-expressing non-viral hyperactive Sleeping Beauty transposon system delivered in a form free of antibiotic resistance marker miniplasmids. The engineered retinal and iris pigment epithelium cells secrete high (141 ± 13 and 222 ± 14 ng PEDF levels in 72 hr in vitro. In vivo studies showed cell survival and insert expression during at least 4 months. Transplantation of the engineered cells to the subretinal space of a rat model of choroidal neovascularization reduces almost 50% of the development of new vessels.

Long interspersed element-1 (LINE-1: passenger or driver in human neoplasms?

Directory of Open Access Journals (Sweden)

Nemanja Rodić

2013-03-01

Full Text Available LINE-1 (L1 retrotransposons make up a significant portion of human genomes, with an estimated 500,000 copies per genome. Like other retrotransposons, L1 retrotransposons propagate through RNA sequences that are reverse transcribed into DNA sequences, which are integrated into new genomic loci. L1 somatic insertions have the potential to disrupt the transcriptome by inserting into or nearby genes. By mutating genes and playing a role in epigenetic dysregulation, L1 transposons may contribute to tumorigenesis. Studies of the "mobilome" have lagged behind other tumor characterizations at the sequence, transcript, and epigenetic levels. Here, we consider evidence that L1 retrotransposons may sometimes drive human tumorigenesis.

Analysis of transposons and repeat composition of the sunflower (Helianthus annuus L.) genome.

Science.gov (United States)

Cavallini, Andrea; Natali, Lucia; Zuccolo, Andrea; Giordani, Tommaso; Jurman, Irena; Ferrillo, Veronica; Vitacolonna, Nicola; Sarri, Vania; Cattonaro, Federica; Ceccarelli, Marilena; Cionini, Pier Giorgio; Morgante, Michele

2010-02-01

A sample-sequencing strategy combined with slot-blot hybridization and FISH was used to study the composition of the repetitive component of the sunflower genome. One thousand six hundred thirty-eight sequences for a total of 954,517 bp were analyzed. The fraction of sequences that can be classified as repetitive using computational and hybridization approaches amounts to 62% in total. Almost two thirds remain as yet uncharacterized in nature. Of those characterized, most belong to the gypsy superfamily of LTR-retrotransposons. Unlike in other species, where single families can account for large fractions of the genome, it appears that no transposon family has been amplified to very high levels in sunflower. All other known classes of transposable elements were also found. One family of unknown nature (contig 61) was the most repeated in the sunflower genome. The evolution of the repetitive component in the Helianthus genus and in other Asteraceae was studied by comparative analysis of the hybridization of total genomic DNAs from these species to the sunflower small-insert library and compared to gene-based phylogeny. Very little similarity is observed between Helianthus species and two related Asteraceae species outside of the genus. Most repetitive elements are similar in annual and perennial Helianthus species indicating that sequence amplification largely predates such divergence. Gypsy-like elements are more represented in the annuals than in the perennials, while copia-like elements are similarly represented, attesting a different amplification history of the two superfamilies of LTR-retrotransposons in the Helianthus genus.

77 FR 17033 - Taking and Importing Marine Mammals: Taking Marine Mammals Incidental to Navy's Training...

Science.gov (United States)

2012-03-23

... take marine mammals by harassment incidental to its training activities at the Gulf of Mexico (GOMEX... Importing Marine Mammals: Taking Marine Mammals Incidental to Navy's Training Activities at the Gulf of Mexico Range Complex AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric...

Marine biodiversity and fishery sustainability.

Science.gov (United States)

Shao, Kwang-Tsao

2009-01-01

Marine fish is one of the most important sources of animal protein for human use, especially in developing countries with coastlines. Marine fishery is also an important industry in many countries. Fifty years ago, many people believed that the ocean was so vast and so resilient that there was no way the marine environment could be changed, nor could marine fishery resources be depleted. Half a century later, we all agree that the depletion of fishery resources is happening mainly due to anthropogenic factors such as overfishing, habitat destruction, pollution, invasive species introduction, and climate change. Since overfishing can cause chain reactions that decrease marine biodiversity drastically, there will be no seafood left after 40 years if we take no action. The most effective ways to reverse this downward trend and restore fishery resources are to promote fishery conservation, establish marine-protected areas, adopt ecosystem-based management, and implement a "precautionary principle." Additionally, enhancing public awareness of marine conservation, which includes eco-labeling, fishery ban or enclosure, slow fishing, and MPA (marine protected areas) enforcement is important and effective. In this paper, we use Taiwan as an example to discuss the problems facing marine biodiversity and sustainable fisheries.

Marine electrical practice

CERN Document Server

Watson, G O

1991-01-01

Marine Engineering Series: Marine Electrical Practice, Sixth Edition focuses on changes in the marine industry, including the application of programmable electronic systems, generators, and motors. The publication first ponders on insulation and temperature ratings of equipment, protection and discrimination, and AC generators. Discussions focus on construction, shaft-drive generators, effect of unbalanced loading, subtransient and transient reactance, protection discrimination, fault current, measurement of ambient air temperature, and basis of machine ratings. The text then examines AC switc

Mariners Weather Log

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Mariners Weather Log (MWL) is a publication containing articles, news and information about marine weather events and phenomena, worldwide environmental impact...

Marine Genomics: A clearing-house for genomic and transcriptomic data of marine organisms

Directory of Open Access Journals (Sweden)

Trent Harold F

2005-03-01

Full Text Available Abstract Background The Marine Genomics project is a functional genomics initiative developed to provide a pipeline for the curation of Expressed Sequence Tags (ESTs and gene expression microarray data for marine organisms. It provides a unique clearing-house for marine specific EST and microarray data and is currently available at http://www.marinegenomics.org. Description The Marine Genomics pipeline automates the processing, maintenance, storage and analysis of EST and microarray data for an increasing number of marine species. It currently contains 19 species databases (over 46,000 EST sequences that are maintained by registered users from local and remote locations in Europe and South America in addition to the USA. A collection of analysis tools are implemented. These include a pipeline upload tool for EST FASTA file, sequence trace file and microarray data, an annotative text search, automated sequence trimming, sequence quality control (QA/QC editing, sequence BLAST capabilities and a tool for interactive submission to GenBank. Another feature of this resource is the integration with a scientific computing analysis environment implemented by MATLAB. Conclusion The conglomeration of multiple marine organisms with integrated analysis tools enables users to focus on the comprehensive descriptions of transcriptomic responses to typical marine stresses. This cross species data comparison and integration enables users to contain their research within a marine-oriented data management and analysis environment.

Sorption kinetics of TNT and RDX in anaerobic freshwater and marine sediments: Batch studies.

Science.gov (United States)

Ariyarathna, Thivanka; Vlahos, Penny; Tobias, Craig; Smith, Richard

2016-01-01

Examination of the partitioning of explosives onto sediment in marine environments is critical to predict the toxicological impacts of worldwide explosive-contaminated sites adjacent to estuaries, wetlands, and the coastal ocean. Marine sediments have been identified as sites of enhanced munitions removal, yet most studies addressing these interactions focus on soils and freshwater sediments. The present study measured the kinetics of 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) sorption onto 2 marine sediments of varying grain sizes (silt vs sand) and organic carbon (OC) content. Abiotic sediment sorption tests were performed at 23 °C, 15 °C, and 4 °C by spiking TNT and RDX solutions directly into anaerobic sediment slurries. Marine sediments showed significantly higher compound uptake rates (0.30-0.80 h(-1) ) than freshwater silt (0.0046-0.0065 h(-1) ) for both compounds, probably because of lower compound solubilities and a higher pH in marine systems. Equilibrium partition constants are on the same order of magnitude for marine silt (1.1-2.0 L kg(-1) sediment) and freshwater silt (1.4-3.1 L kg(-1) sediment) but lower for marine sand (0.72-0.92 L kg(-1) sediment). Total organic carbon content in marine sediments varied linearly with equilibrium partition constants for TNT and was moderately linear for RDX. Uptake rates and equilibrium constants of explosives are inversely correlated to temperature regardless of sediment type because of kinetic barriers associated with low temperatures. © 2015 SETAC.

76 FR 76949 - Marine Mammals

Science.gov (United States)

2011-12-09

...-XR52 Marine Mammals AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric.... 14534 is requested under the authority of the Marine Mammal Protection Act of 1972, as amended (16 U.S.C. 1361 et seq.), the regulations governing the taking and importing of marine mammals (50 CFR part 216...

77 FR 14352 - Marine Mammals

Science.gov (United States)

2012-03-09

...-XB065 Marine Mammals AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric... Marine Mammal Protection Act of 1972, as amended (16 U.S.C. 1361 et seq.), the regulations governing the taking and importing of marine mammals (50 CFR part 216), the Endangered Species Act of 1973, as amended...

75 FR 77616 - Marine Mammals

Science.gov (United States)

2010-12-13

.... 14334] Marine Mammals AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric... Marine Mammal Protection Act of 1972, as amended (16 U.S.C. 1361 et seq.), the regulations governing the taking and importing of marine mammals (50 CFR part 216), the Endangered Species Act of 1973, as amended...

MarineCadastre.gov

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — MarineCadastre.gov is a marine information system that provides authoritative ocean data, offshore planning tools, and technical support to the offshore renewable...

Marine actinobacteria associated with marine organisms and their potentials in producing pharmaceutical natural products.

Science.gov (United States)

Valliappan, Karuppiah; Sun, Wei; Li, Zhiyong

2014-09-01

Actinobacteria are ubiquitous in the marine environment, playing an important ecological role in the recycling of refractory biomaterials and producing novel natural products with pharmic applications. Actinobacteria have been detected or isolated from the marine creatures such as sponges, corals, mollusks, ascidians, seaweeds, and seagrass. Marine organism-associated actinobacterial 16S rRNA gene sequences, i.e., 3,003 sequences, deposited in the NCBI database clearly revealed enormous numbers of actinobacteria associated with marine organisms. For example, RDP classification of these sequences showed that 112 and 62 actinobacterial genera were associated with the sponges and corals, respectively. In most cases, it is expected that these actinobacteria protect the host against pathogens by producing bioactive compounds. Natural products investigation and functional gene screening of the actinobacteria associated with the marine organisms revealed that they can synthesize numerous natural products including polyketides, isoprenoids, phenazines, peptides, indolocarbazoles, sterols, and others. These compounds showed anticancer, antimicrobial, antiparasitic, neurological, antioxidant, and anti-HIV activities. Therefore, marine organism-associated actinobacteria represent an important resource for marine drugs. It is an upcoming field of research to search for novel actinobacteria and pharmaceutical natural products from actinobacteria associated with the marine organisms. In this review, we attempt to summarize the present knowledge on the diversity and natural products production of actinobacteria associated with the marine organisms, based on the publications from 1991 to 2013.

Marine proteomics: a critical assessment of an emerging technology.

Science.gov (United States)

Slattery, Marc; Ankisetty, Sridevi; Corrales, Jone; Marsh-Hunkin, K Erica; Gochfeld, Deborah J; Willett, Kristine L; Rimoldi, John M

2012-10-26

The application of proteomics to marine sciences has increased in recent years because the proteome represents the interface between genotypic and phenotypic variability and, thus, corresponds to the broadest possible biomarker for eco-physiological responses and adaptations. Likewise, proteomics can provide important functional information regarding biosynthetic pathways, as well as insights into mechanism of action, of novel marine natural products. The goal of this review is to (1) explore the application of proteomics methodologies to marine systems, (2) assess the technical approaches that have been used, and (3) evaluate the pros and cons of this proteomic research, with the intent of providing a critical analysis of its future roles in marine sciences. To date, proteomics techniques have been utilized to investigate marine microbe, plant, invertebrate, and vertebrate physiology, developmental biology, seafood safety, susceptibility to disease, and responses to environmental change. However, marine proteomics studies often suffer from poor experimental design, sample processing/optimization difficulties, and data analysis/interpretation issues. Moreover, a major limitation is the lack of available annotated genomes and proteomes for most marine organisms, including several "model species". Even with these challenges in mind, there is no doubt that marine proteomics is a rapidly expanding and powerful integrative molecular research tool from which our knowledge of the marine environment, and the natural products from this resource, will be significantly expanded.

Pigments influence the tolerance of Pseudomonas aeruginosa PAO1 to photodynamically induced oxidative stress

DEFF Research Database (Denmark)

Orlandi, Viviana T; Bolognese, Fabrizio; Chiodaroli, Luca

2015-01-01

by exogenous photosensitizers and visible light. To evaluate whether P. aeruginosa pigments can contribute to its relative tolerance to PDT, we analysed the response to this treatment of isogenic transposon mutants of P. aeruginosa PAO1 with altered pigmentation. In general, in the presence of pigments...

76 FR 41463 - Takes of Marine Mammals Incidental to Specified Activities; Taking Marine Mammals Incidental to a...

Science.gov (United States)

2011-07-14

... requirements, many marine animals may need to remain in areas where they are exposed to chronic stimuli... Marine Mammals Incidental to Specified Activities; Taking Marine Mammals Incidental to a Marine... Atmospheric Administration (NOAA), National Marine Fisheries Service (NMFS). ACTION: Notice; proposed...

[Analysis on sustainable development of marine economy in Jiangsu Province based on marine ecological footprint correction model].

Science.gov (United States)

Yang, Shan; Wang, Yu-ting

2011-03-01

Based on the theories and methods of ecological footprint, the concept of marine ecological footprint was proposed. According to the characteristics of marine environment in Jiangsu Province, five sub-models of marine ecological footprints, including fishery, transporation, marine engineering construction, marine energy, and tidal flat, were constructed. The equilibrium factors of the five marine types were determined by using improved entropy method, and the marine footprints and capacities in Jiangsu Province from 2000 to 2008 were calculated and analyzed. In 2000-2008, the marine ecology footprint per capita in Jiangsu Province increased nearly seven times, from 36.90 hm2 to 252.94 hm2, and the ecological capacity per capita grew steadily, from 105.01 hm2 to 185.49 hm2. In 2000, the marine environment in the Province was in a state of ecological surplus, and the marine economy was in a weak sustainable development state. Since 2004, the marine ecological environment deteriorated sharply, with ecological deficit up to 109660.5 hm2, and the sustainability of marine economy declined. The high ecological footprint of fishery was the main reason for the ecological deficit. Tidal flat was the important reserve resource for the sustainable development of marine economy in Jiangsu Province.

Marine Education Knowledge Inventory.

Science.gov (United States)

Hounshell, Paul B.; Hampton, Carolyn

This 35-item, multiple-choice Marine Education Knowledge Inventory was developed for use in upper elementary/middle schools to measure a student's knowledge of marine science. Content of test items is drawn from oceanography, ecology, earth science, navigation, and the biological sciences (focusing on marine animals). Steps in the construction of…

Factors controlling the compositional variations among the marine and non-marine black shales from Egypt

Energy Technology Data Exchange (ETDEWEB)

Baioumy, Hassan M. [Central Metallurgical R and D Institute, PO Box 87 Helwan, Cairo (Egypt); Ismael, Ismael S. [Faculty of Science, Suez Canal University, Suez (Egypt)

2010-07-01

Non-marine (Jurassic) and marine (Cretaceous) black shales from Egypt were subjected to mineralogical and geochemical analyses to examine the controlling factors of their compositional variations. Non-marine black shales are composed of kaolinite and quartz with traces of gypsum, illite, calcite, feldspars, and dolomite, while marine black shales from the Red Sea area are composed of smectite, kaolinite, quartz, calcite, and dolomite with traces of feldspars. Abu Tartur marine black shales are composed of smectite and quartz with traces of feldspars and gypsum. Non-marine black shales show considerably higher Nb, Ta, Hf, and Zr contents and Th/Yb ratios compared to the marine black shales. On the other hand, marine black shales show considerably higher Cr, V, and Zn contents with positive correlations between these elements and organic carbon (C{sub org.}){sub .} Red Sea black shales have higher Ni/Co, V/Cr, and U/Al ratios. Chondrite normalized values of the medium and heavy rare earth elements (MREEs and HREEs, respectively) are higher in the non-marine black shales compared to the marine black shales. Pyrite from non-marine black shales is characterized by high positive {delta}{sup 34}S isotope values (average of + 9.3 permille). Pyrite from Red Sea black shales has low negative {delta}{sup 34}S values (average of -16.7 permille), pyrite from black shales of the lower member of the Duwi Formation has positive {delta}{sup 34}S values (average of 5.8 permille), while pyrite from marine black shales of the middle member has negative {delta}{sup 34}S values (average of -0.83 permille). Source area composition, weathering conditions, depositional environments, and type of organic matter are considered to be the probable controlling factors of these variations. The more felsic constituents in the source area of non-marine black shales is responsible for the relatively high Nb, Ta, Hf, and Zr contents and Th/Yb ratio. Relatively high kaolinite contents and Chemical

78 FR 30870 - Nomination of Existing Marine Protected Areas to the National System of Marine Protected Areas

Science.gov (United States)

2013-05-23

... Marine Protected Areas to the National System of Marine Protected Areas AGENCY: National Marine Protected...) invited federal, state, commonwealth, and territorial marine protected area (MPA) programs with... of Marine Protected Areas of the United States (Framework), developed in response to Executive Order...

Plastics in the Marine Environment.

Science.gov (United States)

Law, Kara Lavender

2017-01-03

Plastics contamination in the marine environment was first reported nearly 50 years ago, less than two decades after the rise of commercial plastics production, when less than 50 million metric tons were produced per year. In 2014, global plastics production surpassed 300 million metric tons per year. Plastic debris has been detected worldwide in all major marine habitats, in sizes from microns to meters. In response, concerns about risks to marine wildlife upon exposure to the varied forms of plastic debris have increased, stimulating new research into the extent and consequences of plastics contamination in the marine environment. Here, I present a framework to evaluate the current understanding of the sources, distribution, fate, and impacts of marine plastics. Despite remaining knowledge gaps in mass budgeting and challenges in investigating ecological impacts, the increasing evidence of the ubiquity of plastics contamination in the marine environment, the continued rapid growth in plastics production, and the evidence-albeit limited-of demonstrated impacts to marine wildlife support immediate implementation of source-reducing measures to decrease the potential risks of plastics in the marine ecosystem.

Plastics in the Marine Environment

Science.gov (United States)

Law, Kara Lavender

2017-01-01

Plastics contamination in the marine environment was first reported nearly 50 years ago, less than two decades after the rise of commercial plastics production, when less than 50 million metric tons were produced per year. In 2014, global plastics production surpassed 300 million metric tons per year. Plastic debris has been detected worldwide in all major marine habitats, in sizes from microns to meters. In response, concerns about risks to marine wildlife upon exposure to the varied forms of plastic debris have increased, stimulating new research into the extent and consequences of plastics contamination in the marine environment. Here, I present a framework to evaluate the current understanding of the sources, distribution, fate, and impacts of marine plastics. Despite remaining knowledge gaps in mass budgeting and challenges in investigating ecological impacts, the increasing evidence of the ubiquity of plastics contamination in the marine environment, the continued rapid growth in plastics production, and the evidence—albeit limited—of demonstrated impacts to marine wildlife support immediate implementation of source-reducing measures to decrease the potential risks of plastics in the marine ecosystem.

Biosurfactants from marine microorganisms

Directory of Open Access Journals (Sweden)

Suppasil Maneerat

2005-11-01

Full Text Available Biosurfactants are the surface-active molecules synthesized by microorganisms. With the advantage of environmental compatibility, the demand for biosurfactants has been steadily increasing and may eventually replace their chemically synthesized counterparts. Marine biosurfactants produced by some marine microorganisms have been paid more attention, particularly for the bioremediation of the sea polluted by crude oil. This review describes screening of biosurfactant-producing microorganisms, the determination of biosurfactant activity as well as the recovery of marine surfactant. The uses of marine biosurfactants for bioremediation are also discussed.

15 CFR Appendix B to Subpart G of... - Marine Reserve Boundaries

Science.gov (United States)

2010-01-01

....] B.1. Richardson Rock (San Miguel Island) Marine Reserve The Richardson Rock Marine Reserve (Richardson Rock) boundary is defined by the 3 nmi State boundary, the coordinates provided in Table B-1, and the following textual description. The Richardson Rock boundary extends from Point 1 to Point 2 along...

Efficient ensemble forecasting of marine ecology with clustered 1D models and statistical lateral exchange: application to the Red Sea

KAUST Repository

Dreano, Denis

2017-05-24

Forecasting the state of large marine ecosystems is important for many economic and public health applications. However, advanced three-dimensional (3D) ecosystem models, such as the European Regional Seas Ecosystem Model (ERSEM), are computationally expensive, especially when implemented within an ensemble data assimilation system requiring several parallel integrations. As an alternative to 3D ecological forecasting systems, we propose to implement a set of regional one-dimensional (1D) water-column ecological models that run at a fraction of the computational cost. The 1D model domains are determined using a Gaussian mixture model (GMM)-based clustering method and satellite chlorophyll-a (Chl-a) data. Regionally averaged Chl-a data is assimilated into the 1D models using the singular evolutive interpolated Kalman (SEIK) filter. To laterally exchange information between subregions and improve the forecasting skills, we introduce a new correction step to the assimilation scheme, in which we assimilate a statistical forecast of future Chl-a observations based on information from neighbouring regions. We apply this approach to the Red Sea and show that the assimilative 1D ecological models can forecast surface Chl-a concentration with high accuracy. The statistical assimilation step further improves the forecasting skill by as much as 50%. This general approach of clustering large marine areas and running several interacting 1D ecological models is very flexible. It allows many combinations of clustering, filtering and regression technics to be used and can be applied to build efficient forecasting systems in other large marine ecosystems.

Efficient ensemble forecasting of marine ecology with clustered 1D models and statistical lateral exchange: application to the Red Sea

KAUST Repository

Dreano, Denis; Tsiaras, Kostas; Triantafyllou, George; Hoteit, Ibrahim

2017-01-01

Forecasting the state of large marine ecosystems is important for many economic and public health applications. However, advanced three-dimensional (3D) ecosystem models, such as the European Regional Seas Ecosystem Model (ERSEM), are computationally expensive, especially when implemented within an ensemble data assimilation system requiring several parallel integrations. As an alternative to 3D ecological forecasting systems, we propose to implement a set of regional one-dimensional (1D) water-column ecological models that run at a fraction of the computational cost. The 1D model domains are determined using a Gaussian mixture model (GMM)-based clustering method and satellite chlorophyll-a (Chl-a) data. Regionally averaged Chl-a data is assimilated into the 1D models using the singular evolutive interpolated Kalman (SEIK) filter. To laterally exchange information between subregions and improve the forecasting skills, we introduce a new correction step to the assimilation scheme, in which we assimilate a statistical forecast of future Chl-a observations based on information from neighbouring regions. We apply this approach to the Red Sea and show that the assimilative 1D ecological models can forecast surface Chl-a concentration with high accuracy. The statistical assimilation step further improves the forecasting skill by as much as 50%. This general approach of clustering large marine areas and running several interacting 1D ecological models is very flexible. It allows many combinations of clustering, filtering and regression technics to be used and can be applied to build efficient forecasting systems in other large marine ecosystems.

Resistance may be an important mechanism by which marine microbes respond to environmental toxicants*1

Science.gov (United States)

Capriulo, Gerard M.; Flanzenbaum, Jeffrey; Wurster, Charles F.; Rowland, R. George

1983-11-01

The hypothesis, that at least certain marine microbial organisms respond to toxic stress by the development of resistance, was tested using the hypotric marine ciliate Euplotes vannus Muller as the test organism. Resistance to polychlorinated biphenyls (PCB, Aroclor 1254) was developed in E. vannus by exposing the animals to progressively higher PCB concentrations during a period of several months. The resistance to PCB persisted for at least 80 days (greater than 40 generations) after final exposure. This suggests either that genetic selection or persistent (lasting over many cell division cycles) phenotypic trait modification, possibly in the form of Dauermodification, had occurred. If resistance were widespread among marine microbial organisms in polluted environments it would be an important consideration in evaluating the long-term biological impact of both natural and man-induced chemical stress.

75 FR 54095 - Takes of Marine Mammals Incidental to Specified Activities; Low-Energy Marine Seismic Survey in...

Science.gov (United States)

2010-09-03

... Marine Mammals Incidental to Specified Activities; Low- Energy Marine Seismic Survey in the Eastern... low-energy marine seismic survey. Pursuant to the Marine Mammal Protection Act (MMPA), NMFS is... funding provided by the National Science Foundation, a low-energy marine seismic survey. NMFS reviewed SIO...

Linking Marine Ecosystem Services to the North Sea’s Energy Fields in Transnational Marine Spatial Planning

Directory of Open Access Journals (Sweden)

Christina Vogel

2018-06-01

Full Text Available Marine spatial planning temporally and spatially allocates marine resources to different users. The ecosystem approach aims at optimising the social and economic benefits people derive from marine resources while preserving the ecosystem’s health. Marine ecosystem services are defined as the benefits people obtain from marine ecosystems. The aim of this study is to determine which interrelations between marine ecosystem services and the marine energy industry can be identified for use in transnational marine spatial planning exemplified in the North Sea region. As the North Sea is one of the busiest seas worldwide, the risk of impairing the ecosystems through anthropogenic pressures is high. Drawing on a literature-based review, 23 marine ecosystem services provided by the North Sea region were defined and linked to seven offshore energy fields comprising oil and natural gas, wind, tides and currents, waves, salinity gradients, algal biomass, and geothermal heat. The interactions were divided into four categories: dependence, impact, bidirectional, or no interaction. Oil and natural gas, as well as algae biomass, are the fields with the most relations with marine ecosystem services while waves and salinity gradients exhibit the least. Some marine ecosystem services (Conditions for Infrastructure, Regulation of Water Flows, and Cognitive Development are needed for all fields; Recreation and Tourism, Aesthetic and Cultural Perceptions and Traditions, Cognitive Development, and Sea Scape are impacted by all fields. The results of this research provide an improved basis for an ecosystem approach in transnational marine spatial planning.

Drosophila transposon insertions as unknowns for structured inquiry recombination mapping exercises in an undergraduate genetics course.

Science.gov (United States)

Marcus, Jeffrey M; Hughes, Tia M

2009-06-01

Structured inquiry approaches, in which students receive a Drosophila strain of unknown genotype to analyze and map the constituent mutations, are a common feature of many genetics teaching laboratories. The required crosses frustrate many students because they are aware that they are participating in a fundamentally trivial exercise, as the map locations of the genes are already established and have been recalculated thousands of times by generations of students. We modified the traditional structured inquiry approach to include a novel research experience for the students in our undergraduate genetics laboratories. Students conducted crosses with Drosophila strains carrying P[lacW] transposon insertions in genes without documented recombination map positions, representing a large number of unique, but equivalent genetic unknowns. Using the eye color phenotypes associated with the inserts as visible markers, it is straightforward to calculate recombination map positions for the interrupted loci. Collectively, our students mapped 95 genetic loci on chromosomes 2 and 3. In most cases, the calculated 95% confidence interval for meiotic map location overlapped with the predicted map position based on cytology. The research experience evoked positive student responses and helped students better understand the nature of scientific research for little additional cost or instructor effort.

Whale Multi-Disciplinary Studies: A Marine Education Infusion Unit. Northern New England Marine Education Project.

Science.gov (United States)

Maine Univ., Orono. Coll. of Education.

This multidisciplinary unit deals with whales, whaling lore and history, and the interaction of the whale with the complex marine ecosystem. It seeks to teach adaptation of marine organisms. It portrays the concept that man is part of the marine ecosystem and man's activities can deplete and degrade marine ecosystems, endangering the survival of…

A review on marine based nanoparticles and their potential ...

African Journals Online (AJOL)

Infonet

2015-05-06

May 6, 2015 ... potential applications (Table 1) and current information about research on ... easily available source for nanoparticle synthesis with broad variability of ... of marine ecosystem and characterization of marine plants are extremely different ... to develop non-toxic and environment friendly methods to synthesize ...

Supermarket Marine Biology.

Science.gov (United States)

Colby, Jennifer A.; And Others

1995-01-01

Describes a survey used to determine the availability of intact marine vertebrates and live invertebrates in supermarkets. Results shows that local supermarkets frequently provide a variety of intact marine organisms suitable for demonstrations, experiments, or dissections. (ZWH)

Mariner 9 View of Arsia Silva

Science.gov (United States)

1971-01-01

Mariner 9 took this picture of Mars during the closing hours of its approach to the planet on November 13, 1971. The picture has been computer-enhanced with electronic high-pass filtering. The crater-like object at the lower left is about 124 miles (200 kilometers) across and is the same dark spot seen earlier in more distant views. It can be identified on a Mars map as Arsia Silva. The streaks pointing north--more than 1000 kilometers long--are either atmospheric turbulence patterns or dunes formed downwind of the crater. If they are dunes, they are as extensive as the largest in the Sahara in North Africa and those in Peru, South America. The picture was taken from a distance of 65,000 miles about eight hours before Mariner 9 went into orbit around Mars. It was transmitted back to Earth at 10:00 p.m. during the first orbit.Mariner 9 was the first spacecraft to orbit another planet. The spacecraft was designed to continue the atmospheric studies begun by Mariners 6 and 7, and to map over 70% of the Martian surface from the lowest altitude (1500 kilometers [900 miles])and at the highest resolutions (1 kilometer per pixel to 100 meters per pixel) of any previous Mars mission.Mariner 9 was launched on May 30, 1971 and arrived on November 14, 1971.

76 FR 77782 - Takes of Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the...

Science.gov (United States)

2011-12-14

..., 1963), but because of ecological or physiological requirements, many marine animals may need to remain... Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the Commonwealth of the Northern Mariana Islands, February to March 2012 AGENCY: National Marine Fisheries Service (NMFS), National...

78 FR 17359 - Takes of Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the...

Science.gov (United States)

2013-03-21

..., 1963), but because of ecological or physiological requirements, many marine animals may need to remain... Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the Northeast Atlantic Ocean, June to July, 2013 AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...

76 FR 33246 - Takes of Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the...

Science.gov (United States)

2011-06-08

... ecological or physiological requirements, many marine animals may need to remain in areas where they are... Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the Central-Western Bering Sea, August 2011 AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...

75 FR 8652 - Incidental Takes of Marine Mammals During Specified Activities; Marine Geophysical Survey in the...

Science.gov (United States)

2010-02-25

... for marine animals before and during airgun operations. NMFS believes that the realistic possibility... Takes of Marine Mammals During Specified Activities; Marine Geophysical Survey in the Commonwealth of the Northern Mariana Islands, April to June 2010 AGENCY: National Marine Fisheries Service (NMFS...

77 FR 19242 - Takes of Marine Mammals Incidental to Specified Activities; Low-Energy Marine Geophysical Survey...

Science.gov (United States)

2012-03-30

...., 1995; Thorpe, 1963), but because of ecological or physiological requirements, many marine animals may... Marine Mammals Incidental to Specified Activities; Low- Energy Marine Geophysical Survey in the Central Pacific Ocean, May Through June, 2012 AGENCY: National Marine Fisheries Service (NMFS), National Oceanic...

77 FR 25966 - Takes of Marine Mammals Incidental to Specified Activities; Three Marine Geophysical Surveys in...

Science.gov (United States)

2012-05-02

..., 1963), but because of ecological or physiological requirements, many marine animals may need to remain... Marine Mammals Incidental to Specified Activities; Three Marine Geophysical Surveys in the Northeast Pacific Ocean, June Through July 2012 AGENCY: National Marine Fisheries Service, National Oceanic and...

Emerging biopharmaceuticals from marine actinobacteria.

Science.gov (United States)

Hassan, Syed Shams Ul; Anjum, Komal; Abbas, Syed Qamar; Akhter, Najeeb; Shagufta, Bibi Ibtesam; Shah, Sayed Asmat Ali; Tasneem, Umber

2017-01-01

Actinobacteria are quotidian microorganisms in the marine world, playing a crucial ecological role in the recycling of refractory biomaterials and producing novel secondary metabolites with pharmaceutical applications. Actinobacteria have been isolated from the huge area of marine organisms including sponges, tunicates, corals, mollusks, crabs, mangroves and seaweeds. Natural products investigation of the marine actinobacteria revealed that they can synthesize numerous natural products including alkaloids, polyketides, peptides, isoprenoids, phenazines, sterols, and others. These natural products have a potential to provide future drugs against crucial diseases like cancer, HIV, microbial and protozoal infections and severe inflammations. Therefore, marine actinobacteria portray as a pivotal resource for marine drugs. It is an upcoming field of research to probe a novel and pharmaceutically important secondary metabolites from marine actinobacteria. In this review, we attempt to summarize the present knowledge on the diversity, chemistry and mechanism of action of marine actinobacteria-derived secondary metabolites from 2007 to 2016. Copyright © 2016 Elsevier B.V. All rights reserved.

Multisource waveform inversion of marine streamer data using normalized wavefield

KAUST Repository

Choi, Yun Seok

2013-09-01

Multisource full-waveform inversion based on the L1- and L2-norm objective functions cannot be applied to marine streamer data because it does not take into account the unmatched acquisition geometries between the observed and modeled data. To apply multisource full-waveform inversion to marine streamer data, we construct the L1- and L2-norm objective functions using the normalized wavefield. The new residual seismograms obtained from the L1- and L2-norms using the normalized wavefield mitigate the problem of unmatched acquisition geometries, which enables multisource full-waveform inversion to work with marine streamer data. In the new approaches using the normalized wavefield, we used the back-propagation algorithm based on the adjoint-state technique to efficiently calculate the gradients of the objective functions. Numerical examples showed that multisource full-waveform inversion using the normalized wavefield yields much better convergence for marine streamer data than conventional approaches. © 2013 Society of Exploration Geophysicists.

Marine Mineral Exploration

DEFF Research Database (Denmark)

in EEZ areas are fairly unknown; many areas need detailed mapping and mineral exploration, and the majority of coastal or island states with large EEZ areas have little experience in exploration for marine hard minerals. This book describes the systematic steps in marine mineral exploration....... Such exploration requires knowledge of mineral deposits and models of their formation, of geophysical and geochemical exploration methods, and of data evaluation and interpretation methods. These topics are described in detail by an international group of authors. A short description is also given of marine...

76 FR 68720 - Takes of Marine Mammals Incidental to Specified Activities; Low-Energy Marine Geophysical Survey...

Science.gov (United States)

2011-11-07

... Marine Mammals Incidental to Specified Activities; Low- Energy Marine Geophysical Survey in the Western... conducting a low-energy marine geophysical (i.e., seismic) survey in the western tropical Pacific Ocean... Science Foundation (NSF), and ``Environmental Assessment of a Low-Energy Marine Geophysical Survey by the...

76 FR 18167 - Takes of Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the...

Science.gov (United States)

2011-04-01

..., many marine animals may need to remain in areas where they are exposed to chronic stimuli (Richardson... Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the Central Gulf of Alaska, June, 2011 AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric...

76 FR 6430 - Takes of Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the...

Science.gov (United States)

2011-02-04

..., many marine animals may need to remain in areas where they are exposed to chronic stimuli (Richardson... Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the Pacific Ocean off Costa Rica, April Through May, 2011 AGENCY: National Marine Fisheries Service (NMFS), National Oceanic...

76 FR 57959 - Takes of Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the...

Science.gov (United States)

2011-09-19

..., many marine animals may need to remain in areas where they are exposed to chronic stimuli (Richardson... Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the Central Pacific Ocean, November, 2011 Through January, 2012 AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...

78 FR 33357 - Takes of Marine Mammals Incidental to Specified Activities; Taking Marine Mammals Incidental to...

Science.gov (United States)

2013-06-04

... confidence in these values is unknown. Table 3--Marine Mammal Density Estimates Density Species (animals/km\\2... unintentional taking of marine animals occurring incidental to the shock testing which involved large explosives... Marine Mammals Incidental to Specified Activities; Taking Marine Mammals Incidental to Conducting...

76 FR 7548 - Taking and Importing Marine Mammals; Taking of Marine Mammals Incidental to Conducting Precision...

Science.gov (United States)

2011-02-10

... Importing Marine Mammals; Taking of Marine Mammals Incidental to Conducting Precision Strike Weapons Testing and Training by Eglin Air Force Base in the Gulf of Mexico AGENCY: National Marine Fisheries Service... a Letter of Authorization. SUMMARY: In accordance with provisions of the Marine Mammal Protection...

75 FR 16754 - Taking and Importing Marine Mammals; Taking of Marine Mammals Incidental to Conducting Precision...

Science.gov (United States)

2010-04-02

... Importing Marine Mammals; Taking of Marine Mammals Incidental to Conducting Precision Strike Weapons Testing and Training by Eglin Air Force Base in the Gulf of Mexico AGENCY: National Marine Fisheries Service... a Letter of Authorization. SUMMARY: In accordance with provisions of the Marine Mammal Protection...

Microbial dehalogenation of organohalides in marine and estuarine environments.

Science.gov (United States)

Zanaroli, Giulio; Negroni, Andrea; Häggblom, Max M; Fava, Fabio

2015-06-01

Marine sediments are the ultimate sink and a major entry way into the food chain for many highly halogenated and strongly hydrophobic organic pollutants, such as polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), polybrominated diphenylethers (PBDEs) and 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT). Microbial reductive dehalogenation in anaerobic sediments can transform these contaminants into less toxic and more easily biodegradable products. Although little is still known about the diversity of respiratory dehalogenating bacteria and their catabolic genes in marine habitats, the occurrence of dehalogenation under actual site conditions has been reported. This suggests that the activity of dehalogenating microbes may contribute, if properly stimulated, to the in situ bioremediation of marine and estuarine contaminated sediments. Copyright © 2015 Elsevier Ltd. All rights reserved.

THE TOXIC DINOFLAGELLATE GYMNODINIUM CATENATUM (DINOPHYCEAE) REQUIRES MARINE BACTERIA FOR GROWTH(1).

Science.gov (United States)

Bolch, Christopher J S; Subramanian, Thaila A; Green, David H

2011-10-01

Interactions with the bacterial community are increasingly considered to have a significant influence on marine phytoplankton populations. Here we used a simplified dinoflagellate-bacterium experimental culture model to conclusively demonstrate that the toxic dinoflagellate Gymnodinium catenatum H. W. Graham requires growth-stimulatory marine bacteria for postgermination survival and growth, from the point of resting cyst germination through to vegetative growth at bloom concentrations (10(3)  cells · mL(-1) ). Cysts of G. catenatum were germinated and grown in unibacterial coculture with antibiotic-resistant or antibiotic-sensitive Marinobacter sp. DG879 or Brachybacterium sp., and with mixtures of these two bacteria. Addition of antibiotics to cultures grown with antibiotic-sensitive strains of bacteria resulted in death of the dinoflagellate culture, whereas cultures grown with antibiotic-resistant bacteria survived antibiotic addition and continued to grow beyond the 21 d experiment. Removal of either bacterial type from mixed-bacterial dinoflagellate cultures (using an antibiotic) resulted in cessation of dinoflagellate growth until bacterial concentration recovered to preaddition concentrations, suggesting that the bacterial growth factors are used for dinoflagellate growth or are labile. Examination of published reports of axenic dinoflagellate culture indicate that a requirement for bacteria is not universal among dinoflagellates, but rather that species may vary in their relative reliance on, and relationship with, the bacterial community. The experimental model approach described here solves a number of inherent and logical problems plaguing studies of algal-bacterium interactions and provides a flexible and tractable tool that can be extended to examine bacterial interactions with other phytoplankton species. © 2011 Phycological Society of America.

Identification of Genes Involved in Polysaccharide-Independent Staphylococcus aureus Biofilm Formation

Science.gov (United States)

Boles, Blaise R.; Thoendel, Matthew; Roth, Aleeza J.; Horswill, Alexander R.

2010-01-01

Staphylococcus aureus is a potent biofilm former on host tissue and medical implants, and biofilm growth is a critical virulence determinant for chronic infections. Recent studies suggest that many clinical isolates form polysaccharide-independent biofilms. However, a systematic screen for defective mutants has not been performed to identify factors important for biofilm formation in these strains. We created a library of 14,880 mariner transposon mutants in a S. aureus strain that generates a proteinaceous and extracellular DNA based biofilm matrix. The library was screened for biofilm defects and 31 transposon mutants conferred a reproducible phenotype. In the pool, 16 mutants overproduced extracellular proteases and the protease inhibitor α2-macroglobulin restored biofilm capacity to 13 of these mutants. The other 15 mutants in the pool displayed normal protease levels and had defects in genes involved in autolysis, osmoregulation, or uncharacterized membrane proteins. Two transposon mutants of interest in the GraRS two-component system and a putative inositol monophosphatase were confirmed in a flow cell biofilm model, genetically complemented, and further verified in a community-associated methicillin-resistant S. aureus (CA-MRSA) isolate. Collectively, our screen for biofilm defective mutants identified novel loci involved in S. aureus biofilm formation and underscored the importance of extracellular protease activity and autolysis in biofilm development. PMID:20418950

77 FR 4765 - Takes of Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the...

Science.gov (United States)

2012-01-31

... physiological requirements, many marine animals may need to remain in areas where they are exposed to chronic... readily audible to the animals based on measured received levels and the hearing sensitivity of the marine... Marine Mammals Incidental to Specified Activities; Marine Geophysical Survey in the Northwest Pacific...

A Pliocene marine diatom δ18O record of terrestrial-marine feedbacks and orbitally-paced cryogenic brine formation in the McMurdo Dry Valleys

Science.gov (United States)

Dodd, J. P.; Abbott, T.; Gibbons, J. A.

2017-12-01

Orbital frequencies are well documented in a number of terrestrial and marine climate records throughout the Cenozoic; however, assessing the feedbacks and timing of terrestrial-marine systems on glacial-interglacial timescales is often challenging. This is particularly the case in high-latitude, near-shore environments where traditional proxy records like benthic foraminifera are absent. Here we present oxygen isotope (δ18O and δ17O) values from marine diatom silica in the mid-Pliocene (3.5 - 4.7Ma) section of the AND-1B core from McMurdo Sound, Antarctica. Diatom silica δ18O values range between +28.1 and +36.4‰ VSMOW. Over a range of temperatures (0 to 10°C) that reflect both growth and shallow (fall on a mixing line between marine and meteoric waters, which also supports our cryogenic brine hypothesis. The AND-1B δ18O values have an inverse relationship with the stacked benthic foraminifera δ18O record where lower δ18O values in the AND-1B diatom silica correspond with colder intervals, and we interpret variations in the diatom δ18O values as increased brine flux from the MDV to McMurdo Sound. Currently, subsurface brines in the MDV are hydrologically connected with McMurdo Sound. Density-driven transport of these brines from the MDV to the marine costal environments during the warm mid-Pliocene indicate a potentially overlooked terrestrial source of hypersaline waters. Although the lateral extent of these brines is not known, mixing between the terrestrial cryogenic brines and seawater may represent a significant flux of hypersaline water to the marine environment during warmer-than-present global conditions.

Two new pyrrolo-2-aminoimidazoles from a Myanmarese marine sponge, Clathria prolifera.

Science.gov (United States)

Woo, So-Yeun; Win, Nwet Nwet; Wong, Chin Piow; Ito, Takuya; Hoshino, Shotaro; Ngwe, Hla; Aye, Aung Aung; Han, Nang Mya; Zhang, Huiping; Hayashi, Fumiaki; Abe, Ikuro; Morita, Hiroyuki

2018-06-01

Marine organisms such as marine sponges and soft corals are valuable sources of pharmacologically active secondary metabolites. In our ongoing research on the discovery of new secondary metabolites from marine organisms, two new pyrrolo-2-aminoimidazoles, clathriroles A (1) and B (2), were isolated from the water-soluble portion prepared from the methanol and acetone (2:1) extract of the marine sponge, Clathria prolifera, collected in Myanmar. The chemical structures of the isolated compounds were determined using extensive spectroscopic techniques, including NMR, HRESIMS, IR, and optical rotation, and comparisons with the reported literature. The spectroscopic analyses of 1 and 2 suggested that 1 is an enantiomer of antifungal N-methylmanzacidin C isolated from the marine sponge Axinella brevistyla, whereas 2 is a diastereomer of manzacidin D at C-11 isolated from the marine sponge Astrosclera willeyana. To the best of our knowledge, this is the first report of the isolation of the pyrrolo-2-aminoimidazole compounds from C. prolifera. Furthermore, in contrast to the potency of N-methylmanzacidin C against Saccharomyces cerevisiae, the antifungal assay revealed that 1 and 2 lack any activity against this strain. Thus, these observations may suggest that the absolute configurations at both C-9 and C-11 play an important role in controlling the antifungal activity of this type of compound.

CFD simulations on marine burner flames

DEFF Research Database (Denmark)

Cafaggi, Giovanni; Jensen, Peter Arendt; Glarborg, Peter

The marine industry is changing with new demands concerning high energy efficiency, fuel flexibility and lower emissions of NOX and SOX. A collaboration between the company Alfa Laval and Technical University of Denmark has been established to support the development of the next generation...... of marine burners. The resulting auxiliary boilers shall be compact and able to operate with different fuel types, while reducing NOX emissions. The specific boiler object of this study uses a swirl stabilized liquid fuel burner, with a pressure swirl spill-return atomizer (Fig.1). The combustion chamber...... is enclosed in a water jacket used for water heating and evaporation, and a convective heat exchanger at the furnace outlet super-heats the steam. The purpose of the present study is to gather detailed knowledge about the influence of fuel spray conditions on marine utility boiler flames. The main goal...

Future Retention of the Marine Corps Top Performing Aviators

Science.gov (United States)

2016-06-10

balance, and Aviator Continuation Pay. 15. SUBJECT TERMS USMC Aviator Retention, USMC Aviation Manpower, Marine Corps Talent Management , Marine Corps...Wing Aircraft: Includes AH-1, UH-1, and CH-53 platforms. Talent Management : The process through which employers anticipate and meet their needs for...a bias due to his experience as an executive officer responsible for the talent management of company grade WTI aviators. All attempts have been

Striking structural dynamism and nucleotide sequence variation of the transposon Galileo in the genome of Drosophila mojavensis.

Science.gov (United States)

Marzo, Mar; Bello, Xabier; Puig, Marta; Maside, Xulio; Ruiz, Alfredo

2013-02-04

Galileo is a transposable element responsible for the generation of three chromosomal inversions in natural populations of Drosophila buzzatii. Although the most characteristic feature of Galileo is the long internally-repetitive terminal inverted repeats (TIRs), which resemble the Drosophila Foldback element, its transposase-coding sequence has led to its classification as a member of the P-element superfamily (Class II, subclass 1, TIR order). Furthermore, Galileo has a wide distribution in the genus Drosophila, since it has been found in 6 of the 12 Drosophila sequenced genomes. Among these species, D. mojavensis, the one closest to D. buzzatii, presented the highest diversity in sequence and structure of Galileo elements. In the present work, we carried out a thorough search and annotation of all the Galileo copies present in the D. mojavensis sequenced genome. In our set of 170 Galileo copies we have detected 5 Galileo subfamilies (C, D, E, F, and X) with different structures ranging from nearly complete, to only 2 TIR or solo TIR copies. Finally, we have explored the structural and length variation of the Galileo copies that point out the relatively frequent rearrangements within and between Galileo elements. Different mechanisms responsible for these rearrangements are discussed. Although Galileo is a transposable element with an ancient history in the D. mojavensis genome, our data indicate a recent transpositional activity. Furthermore, the dynamism in sequence and structure, mainly affecting the TIRs, suggests an active exchange of sequences among the copies. This exchange could lead to new subfamilies of the transposon, which could be crucial for the long-term survival of the element in the genome.

Marine medicinal glycomics

Directory of Open Access Journals (Sweden)

Vitor Hugo Pomin

2014-01-01

Full Text Available Glycomics is an international initiative aimed to understand the structure and function of the glycans from a given type of cell, tissue, organism, kingdom or even environment, as found under certain conditions. Glycomics is one of the latest areas of intense biological research. Glycans of marine sources are unique in terms of structure and function. They differ considerably from those of terrestrial origin. This review discusses the most known marine glycans of potential therapeutic properties. They are chitin, chitosan, and sulfated polysaccharides named glycosaminoglycans, sulfated fucans and sulfated galactans. Their medical actions are very broad. When certain structural requirements are found, these glycans can exhibit beneficial effects in inflammation, coagulation, thrombosis, cancer growth/metastasis and vascular biology. Both structure and therapeutic mechanisms of action of these marine glycans are discussed here in straight context with the current glycomic age through a project suggestively named Marine Medicinal Glycomics.

75 FR 19670 - Marine Highway Projects

Science.gov (United States)

2010-04-15

... DEPARTMENT OF TRANSPORTATION Maritime Administration Marine Highway Projects ACTION: Solicitation of applications for Marine highway projects. SUMMARY: The Department of Transportation is soliciting applications for Marine Highway Projects as specified in the America's Marine Highway Program Final Rule, MARAD...

Western Indian Ocean Journal of Marine Science - Vol 9, No 1 (2010)

African Journals Online (AJOL)

Coastal Marine Pollution in Dar es Salaam (Tanzania) relative to Recommended Environmental Quality Targets for the Western Indian Ocean · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. JF Machiwa, 17-30 ...

Marine and Estuarine Pollution.

Science.gov (United States)

Reish, Donald J.

1978-01-01

Presents a literature review of the effects of various pollutants on marine and estuarine organisms, covering publications of 1976-77. This review includes: (1) effects of pesticides, dredging, dumping, sludge, and petroleum hydrocarbons; and (2) diseases and tissue abnormalities. A list of 441 references is also presented. (HM)

Review of scientific information on impacts of seismic sound on fish, invertebrates, marine turtles and marine mammals

Energy Technology Data Exchange (ETDEWEB)

Bain, H.

2004-09-01

This review of scientific literature on impacts of seismic sound on aquatic organisms was initiated following a 2003 workshop to develop a decision framework for seismic survey referrals in Canadian waters. That workshop revealed that there are sources of uncertainty about the effects of seismic sound on aquatic organisms. It was determined that seismic sounds on the marine environment are not completely without consequence nor are they certain to result in serious harm. Following the workshop, and in order to clearly determine the level of risk posed by seismic sounds, teams of scientists prepared reviews of literature on experimental studies and field monitoring on the effects of seismic sound on marine organisms. Standards and mitigation methods were also reviewed. The scientific deliberations resulted in a body of information that allowed several conclusions to be reached that provide a scientific basis for developing a regulatory framework for conducting seismic surveys in marine environments. This paper presented literature highlights regarding: habitat concern; management considerations; physical and behavioural effects on fish; functional uses of sound; physical, physiological and behavioural effects on invertebrates; effects of seismic sound on zooplankton, eggs, larvae of fish and invertebrates; effect of seismic sound on marine turtles; and mortality, physical and behavioural effects on marine mammals. The literature review sought to seek if seismic sound contributed to displacement and migratory diversion; changes in dive and respiratory patterns; changes in social behaviour; and changes in vocalisation patterns. Several areas of future research needs were identified following this literature review which revealed that the long-term effects of seismic sound on marine animals remain inconclusive. 2 refs., 1 fig.

77 FR 841 - Taking and Importing Marine Mammals: Taking Marine Mammals Incidental to U.S. Navy Operations of...

Science.gov (United States)

2012-01-06

... and the Single Ping Equivalent (SPE) To model potential impacts to marine animals from exposure to... Atmospheric Administration 50 CFR Part 218 Taking and Importing Marine Mammals: Taking Marine Mammals... [Docket No. 110808485-1534-01] RIN 0648-BB14 Taking and Importing Marine Mammals: Taking Marine Mammals...

Identifying Sources of Marine Litter

OpenAIRE

VEIGA Joana Mira; FLEET David; KINSEY Sue; NILSSON Per; VLACHOGIANNI Thomais; WERNER Stefanie; GALGANI Francois; THOMPSON Richard; DAGEVOS Jeroen; GAGO Jesus; SOBRAL Paula; CRONIN Richard

2016-01-01

Marine litter is a global problem causing harm to marine wildlife, coastal communities and maritime activities. It also embodies an emerging concern for human health and safety. The reduction of marine litter pollution poses a complex challenge for humankind, requiring adjustments in human behaviour as well as in the different phases of the life-cycle of products and across multiple economic sectors. The Marine Strategy Framework Directive (MSFD) requires European Member States to monitor...

Chemical Investigations of Marine Filamentous and Zoosporic Fungi and Studies in Marine Microbial Chemical Ecology

OpenAIRE

Jenkins, Kelly M.

1998-01-01

The natural products chemistry of marine microorganisms is an emerging area of organic chemistry with the aim of discovering novel secondary metabolites exhibiting both biomedical and ecological activities. While marine bacteria have proven to be a productive source of new natural products, there are many groups of marine microorganisms which have not been fully investigated. In particular, marine fungi represent an untapped and potentially novel source of bioactive secondary metabolites. Whi...

Bioactive benzopyrone derivatives from new recombinant fusant of marine Streptomyces.

Science.gov (United States)

El-Gendy, Mervat M A; Shaaban, M; El-Bondkly, A M; Shaaban, K A

2008-07-01

In our searching program for bioactive secondary metabolites from marine Streptomycetes, three microbial benzopyrone derivatives (1-3), 7-methylcoumarin (1) and two flavonoides, rhamnazin (2) and cirsimaritin (3), were obtained during the working up of the ethyl acetate fraction of a marine Streptomyces fusant obtained from protoplast fusion between Streptomyces strains Merv 1996 and Merv 7409. The structures of the three compounds (1-3) were established by nuclear magnetic resonance, mass, UV spectra, and by comparison with literature data. Marine Streptomyces strains were identified based on their phenotypic and chemotypic characteristics as two different bioactive strains of the genus Streptomyces. We described here the fermentation, isolation, as well as the biological activity of these bioactive compounds. The isolated compounds (1-3) are reported here as microbial products for the first time.

Oceanic processes in marine pollution

International Nuclear Information System (INIS)

Baumgartner, D.J.; Duedall, I.W.

1990-01-01

This book covers the following areas: bioaccumulation of Polycyclic Aromatic hydrocarbons in marine environments; behavior of drilling fluid discharges off the coast of California; effects of drilling fluids on marine organisms; and the effects of radioactive waste disposal on marine amphipods

pBAM1: an all-synthetic genetic tool for analysis and construction of complex bacterial phenotypes

Directory of Open Access Journals (Sweden)

Arévalo-Rodríguez Miguel

2011-02-01

Full Text Available Abstract Background Since publication in 1977 of plasmid pBR322, many breakthroughs in Biology have depended on increasingly sophisticated vector platforms for analysis and engineering of given bacterial strains. Although restriction sites impose a certain format in the procedures for assembling cloned genes, every attempt thus far to standardize vector architecture and nomenclature has ended up in failure. While this state of affairs may still be tolerable for traditional one-at-a-time studies of single genes, the onset of systems and synthetic biology calls for a simplification -along with an optimization- of the currently unwieldy pool of genetic tools. Results The functional DNA sequences present in the natural bacterial transposon Tn5 have been methodically edited and refactored for the production of a multi-purpose genetic tool named pBAM1, which allows a range of manipulations in the genome of Gram-negative bacteria. This all-synthetic construct enhances the power of mini-transposon vectors for either de-construction or re-construction of phenotypes á la carte by incorporating features inspired in systems engineering: modularity, re-usability, minimization, and compatibility with other genetic tools. pBAM1 bears an streamlined, restriction site-freed and narrow-host range replication frame bearing the sequences of R6K oriV, oriT and an ampicillin resistance marker. These go along with a business module that contains a host-independent and hyperactive transposition platform for in vivo or in vitro insertion of desired DNA into the genome of the target bacterium. All functional sequences were standardized for a straightforward replacement by equivalent counterparts, if required. pBAM1 can be delivered into recipient cells by either mating or electroporation, producing transposon insertion frequencies of 1.8 × 10-3 and 1.02 × 10-7, respectively in the soil bacterium Pseudomonas putida. Analyses of the resulting clones revealed a 100% of

Massive Expansion of Marine Archaea During The Early Albian Oceanic Anoxic Event 1B

Science.gov (United States)

Kuypers, M. M.; Kuypers, M. M.; Blokker, P.; Erbacher, J.; Kinkel, H.; Pancost, R. D.; Pancost, R. D.; Schouten, S.; Sinninghe Damsté, J. S.

2001-12-01

Oceanic anoxic events (OAEs), periods of globally enhanced burial of organic matter (OM) in the marine realm, played an important role in the mid-Cretaceous `greenhouse climate' by effectively reducing atmospheric carbon dioxide concentrations. It is generally believed that these OAEs were caused either by decreased remineralisation or increased production of phytoplanktonic OM. Here we show that enhanced organic carbon (OC) burial during the early Albian OAE1b (~112 My) was caused by a different process. Combined biogeochemical and stable carbon isotopic analyses indicate that black shales from this period contain up to 80% of OC derived from archaea. Archaea-derived isoprenoidal tetraether membrane lipids and free and macromolecularly bound isoprenoid alkanes are abundantly present in these black shales. More specifically the presence of certain ether lipids (cyclic biphytane tetraethers) indicates representatives of the pelagic archaea. To the best of our knowledge this is the earliest fossil evidence for marine planktonic archaea, extending their geological record by more than 60 million years. The diversity of archaeal lipids recovered from the OAE1b black shales suggests that they derive from a multitude of archaeal species. However, the specific 13C enrichment of all such lipids indicates a common `heavy' (13C-rich) carbon source for the archaea and/or a common pathway of carbon-fixation with a reduced 13C fractionation effect compared to the Calvin cycle used by algae, cyanobacteria and higher plants. The large differences (up to 12%) in 13C/12C ratios between the algal biomarkers and the much more abundant archaeal molecular fossils suggest that the latter were not living heterotrophically on photoautotrophic biomass. It seems likely that the archaea present during OAE1b used a chemical energy source (possibly ammonium) for carbon fixation since photoautotrophy within the domain of the Archaea is restricted to only a few species from hypersaline

Mercury pollution for marine environment at Farwa Island, Libya.

Science.gov (United States)

Banana, Adel A S; Mohamed, R M S Radin; Al-Gheethi, A A S

2016-01-01

Farwa is an Island in Libya receives petrochemical wastes generated from General Company of Chemical Industries (GCCI) since more than 40 years. The present work aimed to determine the concentrations of mercury (Hg(+2)) in fish, marine plants and sediment collected from Farwa lagoon to evaluate effect of industrial wastewater from GCCI on the marine environment. Hundred and twelve samples of fish, pearl oyster, cuttlefish sediments and marine plants were analyzed to determine Hg(2+) concentration during the period from January to August 2014 by using Atomic Absorption Spectrometer (AAS). The highest concentration of Hg(2+) was detected in Pinctada radiata (11.67 ± 3.30 μgg (-1)) followed by Serranus scriba (6.37 ± 0.11 μg g (-1)) and Epinephelus marginatus (6.19 ± 0.02 μg g (-1)). About 75 % of marine plants contained the maximum contaminations during the summer season. In fish samples Hg(2+) concentrations exceeded the levels provided by international standards. The fish at Farwa lagoon is heavily contaminated with Hg(2+) which may represent a source for mercury poisoning for human.

Analysis of the Pseudoalteromonas tunicata genome reveals properties of a surface-associated life style in the marine environment.

Directory of Open Access Journals (Sweden)

Torsten Thomas

Full Text Available BACKGROUND: Colonisation of sessile eukaryotic host surfaces (e.g. invertebrates and seaweeds by bacteria is common in the marine environment and is expected to create significant inter-species competition and other interactions. The bacterium Pseudoalteromonas tunicata is a successful competitor on marine surfaces owing primarily to its ability to produce a number of inhibitory molecules. As such P. tunicata has become a model organism for the studies into processes of surface colonisation and eukaryotic host-bacteria interactions. METHODOLOGY/PRINCIPAL FINDINGS: To gain a broader understanding into the adaptation to a surface-associated life-style, we have sequenced and analysed the genome of P. tunicata and compared it to the genomes of closely related strains. We found that the P. tunicata genome contains several genes and gene clusters that are involved in the production of inhibitory compounds against surface competitors and secondary colonisers. Features of P. tunicata's oxidative stress response, iron scavenging and nutrient acquisition show that the organism is well adapted to high-density communities on surfaces. Variation of the P. tunicata genome is suggested by several landmarks of genetic rearrangements and mobile genetic elements (e.g. transposons, CRISPRs, phage. Surface attachment is likely to be mediated by curli, novel pili, a number of extracellular polymers and potentially other unexpected cell surface proteins. The P. tunicata genome also shows a utilisation pattern of extracellular polymers that would avoid a degradation of its recognised hosts, while potentially causing detrimental effects on other host types. In addition, the prevalence of recognised virulence genes suggests that P. tunicata has the potential for pathogenic interactions. CONCLUSIONS/SIGNIFICANCE: The genome analysis has revealed several physiological features that would provide P. tunciata with competitive advantage against other members of the surface

Analysis of the Pseudoalteromonas tunicata genome reveals properties of a surface-associated life style in the marine environment.

Science.gov (United States)

Thomas, Torsten; Evans, Flavia F; Schleheck, David; Mai-Prochnow, Anne; Burke, Catherine; Penesyan, Anahit; Dalisay, Doralyn S; Stelzer-Braid, Sacha; Saunders, Neil; Johnson, Justin; Ferriera, Steve; Kjelleberg, Staffan; Egan, Suhelen

2008-09-24

Colonisation of sessile eukaryotic host surfaces (e.g. invertebrates and seaweeds) by bacteria is common in the marine environment and is expected to create significant inter-species competition and other interactions. The bacterium Pseudoalteromonas tunicata is a successful competitor on marine surfaces owing primarily to its ability to produce a number of inhibitory molecules. As such P. tunicata has become a model organism for the studies into processes of surface colonisation and eukaryotic host-bacteria interactions. To gain a broader understanding into the adaptation to a surface-associated life-style, we have sequenced and analysed the genome of P. tunicata and compared it to the genomes of closely related strains. We found that the P. tunicata genome contains several genes and gene clusters that are involved in the production of inhibitory compounds against surface competitors and secondary colonisers. Features of P. tunicata's oxidative stress response, iron scavenging and nutrient acquisition show that the organism is well adapted to high-density communities on surfaces. Variation of the P. tunicata genome is suggested by several landmarks of genetic rearrangements and mobile genetic elements (e.g. transposons, CRISPRs, phage). Surface attachment is likely to be mediated by curli, novel pili, a number of extracellular polymers and potentially other unexpected cell surface proteins. The P. tunicata genome also shows a utilisation pattern of extracellular polymers that would avoid a degradation of its recognised hosts, while potentially causing detrimental effects on other host types. In addition, the prevalence of recognised virulence genes suggests that P. tunicata has the potential for pathogenic interactions. The genome analysis has revealed several physiological features that would provide P. tunciata with competitive advantage against other members of the surface-associated community. We have also identified properties that could mediate interactions

High Performance Marine Vessels

CERN Document Server

Yun, Liang

2012-01-01

High Performance Marine Vessels (HPMVs) range from the Fast Ferries to the latest high speed Navy Craft, including competition power boats and hydroplanes, hydrofoils, hovercraft, catamarans and other multi-hull craft. High Performance Marine Vessels covers the main concepts of HPMVs and discusses historical background, design features, services that have been successful and not so successful, and some sample data of the range of HPMVs to date. Included is a comparison of all HPMVs craft and the differences between them and descriptions of performance (hydrodynamics and aerodynamics). Readers will find a comprehensive overview of the design, development and building of HPMVs. In summary, this book: Focuses on technology at the aero-marine interface Covers the full range of high performance marine vessel concepts Explains the historical development of various HPMVs Discusses ferries, racing and pleasure craft, as well as utility and military missions High Performance Marine Vessels is an ideal book for student...

Hypoxia in the changing marine environment

Science.gov (United States)

Zhang, J.; Cowie, G.; Naqvi, S. W. A.

2013-03-01

The predicted future of the global marine environment, as a combined result of forcing due to climate change (e.g. warming and acidification) and other anthropogenic perturbation (e.g. eutrophication), presents a challenge to the sustainability of ecosystems from tropics to high latitudes. Among the various associated phenomena of ecosystem deterioration, hypoxia can cause serious problems in coastal areas as well as oxygen minimum zones in the open ocean (Diaz and Rosenberg 2008 Science 321 926-9, Stramma et al 2008 Science 320 655-8). The negative impacts of hypoxia include changes in populations of marine organisms, such as large-scale mortality and behavioral responses, as well as variations of species distributions, biodiversity, physiological stress, and other sub-lethal effects (e.g. growth and reproduction). Social and economic activities that are related to services provided by the marine ecosystems, such as tourism and fisheries, can be negatively affected by the aesthetic outcomes as well as perceived or real impacts on seafood quality (STAP 2011 (Washington, DC: Global Environment Facility) p 88). Moreover, low oxygen concentration in marine waters can have considerable feedbacks to other compartments of the Earth system, like the emission of greenhouse gases to the atmosphere, and can affect the global biogeochemical cycles of nutrients and trace elements. It is of critical importance to prediction and adaptation strategies that the key processes of hypoxia in marine environments be precisely determined and understood (cf Zhang et al 2010 Biogeosciences 7 1-24).

76 FR 12070 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2011-03-04

... Energy's EROS operations in 2010: Marine mammals Biological impacts Company Structure Dates sighted... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...

77 FR 45341 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2012-07-31

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...). SUMMARY: In accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations...

77 FR 16539 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2012-03-21

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...). SUMMARY: In accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations...

78 FR 22517 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2013-04-16

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...). SUMMARY: In accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations...

75 FR 31423 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2010-06-03

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...: In accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification...

78 FR 13865 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2013-03-01

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...). SUMMARY: In accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations...

77 FR 39485 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2012-07-03

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and...). SUMMARY: In accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations...

Marine Fish Hybridization

KAUST Repository

He, Song

2017-04-01

Natural hybridization is reproduction (without artificial influence) between two or more species/populations which are distinguishable from each other by heritable characters. Natural hybridizations among marine fishes were highly underappreciated due to limited research effort; it seems that this phenomenon occurs more often than is commonly recognized. As hybridization plays an important role in biodiversity processes in the marine environment, detecting hybridization events and investigating hybridization is important to understand and protect biodiversity. The first chapter sets the framework for this disseration study. The Cohesion Species Concept was selected as the working definition of a species for this study as it can handle marine fish hybridization events. The concept does not require restrictive species boundaries. A general history and background of natural hybridization in marine fishes is reviewed during in chapter as well. Four marine fish hybridization cases were examed and documented in Chapters 2 to 5. In each case study, at least one diagnostic nuclear marker, screened from among ~14 candidate markers, was found to discriminate the putative hybridizing parent species. To further investigate genetic evidence to support the hybrid status for each hybrid offspring in each case, haploweb analysis on diagnostic markers (nuclear and/or mitochondrial) and the DAPC/PCA analysis on microsatellite data were used. By combining the genetic evidences, morphological traits, and ecological observations together, the potential reasons that triggered each hybridization events and the potential genetic/ecology effects could be discussed. In the last chapter, sequences from 82 pairs of hybridizing parents species (for which COI barcoding sequences were available either on GenBank or in our lab) were collected. By comparing the COI fragment p-distance between each hybridizing parent species, some general questions about marine fish hybridization were discussed: Is

77 FR 50289 - Taking and Importing Marine Mammals: Taking Marine Mammals Incidental to U.S. Navy Operations of...

Science.gov (United States)

2012-08-20

... areas of high marine animal densities, we believe that the incidental take of marine mammals would... response, tagging of free-ranging marine animals at-sea, and radar-based detection of marine mammals from... effects of sonar use will not be greater on animals listed under the ESA than the effects on other marine...

Moving through the stressed genome: Emerging regulatory roles for transposons in plant stress tolerance

Directory of Open Access Journals (Sweden)

Negi Pooja

2016-10-01

Full Text Available The recognition of a positive correlation between organism genome size with its transposable element (TE content, represents a key discovery of the field of genome biology. Considerable evidence accumulated since then suggests the involvement of TEs in genome structure, evolution and function. The global genome reorganization brought about by transposon activity might play an adaptive/regulatory role in the host response to environmental challenges, reminiscent of McClintock’s original ’Controlling Element’ hypothesis. This regulatory aspect of TEs is also garnering support in light of the recent evidences which project TEs as distributed genomic control modules. According to this view, TEs are capable of actively reprogramming host genes circuits and ultimately fine-tuning the host response to specific environmental stimuli. Moreover, the stress-induced changes in epigenetic status of TE activity may allow TEs to propagate their stress responsive elements to host genes; the resulting genome fluidity can permit phenotypic plasticity and adaptation to stress. Given their predominating presence in the plant genomes, nested organization in the genic regions and potential regulatory role in stress response, TEs hold unexplored potential for crop improvement programs. This review intends to present the current information about the roles played by TEs in plant genome organization, evolution and function, and highlight the regulatory mechanisms in plant stress responses. We will also briefly discuss the connection between TE activity, host epigenetic response and phenotypic plasticity as a critical link for traversing the translational bridge from a purely basic study of TEs, to the applied field of stress adaptation and crop improvement.

Moving through the Stressed Genome: Emerging Regulatory Roles for Transposons in Plant Stress Response.

Science.gov (United States)

Negi, Pooja; Rai, Archana N; Suprasanna, Penna

2016-01-01

The recognition of a positive correlation between organism genome size with its transposable element (TE) content, represents a key discovery of the field of genome biology. Considerable evidence accumulated since then suggests the involvement of TEs in genome structure, evolution and function. The global genome reorganization brought about by transposon activity might play an adaptive/regulatory role in the host response to environmental challenges, reminiscent of McClintock's original 'Controlling Element' hypothesis. This regulatory aspect of TEs is also garnering support in light of the recent evidences, which project TEs as "distributed genomic control modules." According to this view, TEs are capable of actively reprogramming host genes circuits and ultimately fine-tuning the host response to specific environmental stimuli. Moreover, the stress-induced changes in epigenetic status of TE activity may allow TEs to propagate their stress responsive elements to host genes; the resulting genome fluidity can permit phenotypic plasticity and adaptation to stress. Given their predominating presence in the plant genomes, nested organization in the genic regions and potential regulatory role in stress response, TEs hold unexplored potential for crop improvement programs. This review intends to present the current information about the roles played by TEs in plant genome organization, evolution, and function and highlight the regulatory mechanisms in plant stress responses. We will also briefly discuss the connection between TE activity, host epigenetic response and phenotypic plasticity as a critical link for traversing the translational bridge from a purely basic study of TEs, to the applied field of stress adaptation and crop improvement.

Moving through the Stressed Genome: Emerging Regulatory Roles for Transposons in Plant Stress Response

Science.gov (United States)

Negi, Pooja; Rai, Archana N.; Suprasanna, Penna

2016-01-01

The recognition of a positive correlation between organism genome size with its transposable element (TE) content, represents a key discovery of the field of genome biology. Considerable evidence accumulated since then suggests the involvement of TEs in genome structure, evolution and function. The global genome reorganization brought about by transposon activity might play an adaptive/regulatory role in the host response to environmental challenges, reminiscent of McClintock's original ‘Controlling Element’ hypothesis. This regulatory aspect of TEs is also garnering support in light of the recent evidences, which project TEs as “distributed genomic control modules.” According to this view, TEs are capable of actively reprogramming host genes circuits and ultimately fine-tuning the host response to specific environmental stimuli. Moreover, the stress-induced changes in epigenetic status of TE activity may allow TEs to propagate their stress responsive elements to host genes; the resulting genome fluidity can permit phenotypic plasticity and adaptation to stress. Given their predominating presence in the plant genomes, nested organization in the genic regions and potential regulatory role in stress response, TEs hold unexplored potential for crop improvement programs. This review intends to present the current information about the roles played by TEs in plant genome organization, evolution, and function and highlight the regulatory mechanisms in plant stress responses. We will also briefly discuss the connection between TE activity, host epigenetic response and phenotypic plasticity as a critical link for traversing the translational bridge from a purely basic study of TEs, to the applied field of stress adaptation and crop improvement. PMID:27777577

Molecular diversity of fungi from marine oxygen-deficient environments (ODEs)

Digital Repository Service at National Institute of Oceanography (India)

Manohar, C.S.; Forster, D.; Kauff, F.; Stoeck, T.

. Sparrow Jr F K (1936) Biological observations of the marine fungi of woods hole waters. Biol Bull 70: 236-263. States JS & Christensen M (2001) Fungi Associated with Biological Soil Crusts in Desert Grasslands of Utah and Wyoming. Mycologia 93: 432... version: Biology of marine fungi. Ed. by: Raghukumar, C. (Prog. Mol. Subcellular Biol). Springer, vol.53 (Chap 10); 2012; 189-208 Chapter # 10 Molecular diversity of fungi from marine oxygen-deficient environments (ODEs) Cathrine S. Jebaraj 1...

75 FR 972 - Nomination of Existing Marine Protected Areas to the National System of Marine Protected Areas

Science.gov (United States)

2010-01-07

... Marine Protected Areas to the National System of Marine Protected Areas AGENCY: NOAA, Department of... Federal, State and territorial marine protected area programs to join the National System of Marine Protected Areas. SUMMARY: NOAA and the Department of the Interior (DOI) invited Federal, State, commonwealth...

76 FR 35856 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2011-06-20

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and... accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification is hereby...

75 FR 8921 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2010-02-26

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and... accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification is hereby...

76 FR 33704 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2011-06-09

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and... accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification is hereby...

77 FR 10481 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2012-02-22

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and... accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification is hereby...

76 FR 23570 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2011-04-27

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and... accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification is hereby...

75 FR 28566 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2010-05-21

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and... accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification is hereby...

75 FR 54851 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2010-09-09

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and... accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification is hereby...

75 FR 38078 - Incidental Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal...

Science.gov (United States)

2010-07-01

... Taking of Marine Mammals; Taking of Marine Mammals Incidental to the Explosive Removal of Offshore Structures in the Gulf of Mexico AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and... accordance with the Marine Mammal Protection Act (MMPA) and implementing regulations, notification is hereby...

Speciated mercury at marine, coastal, and inland sites in New England – Part 1: Temporal variability

Directory of Open Access Journals (Sweden)

H. Mao

2012-06-01

Full Text Available A comprehensive analysis was conducted using long-term continuous measurements of gaseous elemental mercury (Hg⁰, reactive gaseous mercury (RGM, and particulate phase mercury (Hg^P at coastal (Thompson Farm, denoted as TF, marine (Appledore Island, denoted as AI, and elevated inland (Pac Monadnock, denoted as PM sites from the AIRMAP Observatories in southern New Hampshire, USA. Decreasing trends in background Hg⁰ were identified in the 7.5- and 5.5-yr records at TF and PM with decline rates of 3.3 parts per quadrillion by volume (ppqv yr⁻¹ and 6.3 ppqv yr⁻¹, respectively. Common characteristics at these sites were the reproducible annual cycle of Hg⁰ with its maximum in winter-spring and minimum in fall, comprised of a positive trend in the warm season (spring – early fall and a negative one in the cool season (late fall – winter. Year-to-year variability was observed in the warm season decline in Hg⁰ at TF varying from a minimum total (complete seasonal loss of 43 ppqv in 2009 to a maximum of 92 ppqv in 2005, whereas variability remained small at AI and PM. The coastal site TF differed from the other two sites with its exceptionally low levels (as low as below 50 ppqv in the nocturnal inversion layer possibly due to dissolution in dew water. Measurements of Hg⁰ at PM exhibited the smallest diurnal to annual variability among the three environments, where peak levels rarely exceeded 250 ppqv and the minimum was typically 100 ppqv. It should be noted that summertime diurnal patterns at TF and AI were opposite in phase indicating strong sink(s for Hg⁰ during the day in the marine boundary layer, which was consistent with the hypothesis of Hg⁰ oxidation by halogen radicals there. Mixing ratios of RGM in the coastal and marine boundary layers reached annual maxima in spring and minima in fall, whereas at PM levels were generally

A Demonstration Marine Biodiversity Observation Network (MBON): Understanding Marine Life and its Role in Maintaining Ecosystem Services

Science.gov (United States)

Muller-Karger, F. E.; Iken, K.; Miller, R. J.; Duffy, J. E.; Chavez, F.; Montes, E.

2016-02-01

The U.S. Federal government (NOAA, NASA, BOEM, and the Smithsonian Institution), academic researchers, and private partners are laying the foundation for a Marine Biodiversity Observation Network (MBON). The goals of the network are to: 1) Observe and understand life, from microbes to whales, in different coastal and continental shelf habitats; 2) Define an efficient set of observations required for implementing a useful MBON; 3) Develop technology for biodiversity assessments including emerging environmental DNA (eDNA), remote sensing, and image analysis methods to coordinate with classical sampling; 4) Integrate and synthesize information in coordination with the Integrated Ocean Observing System (IOOS), the international Group on Earth Observations Biodiversity Observation Network(GEO BON), and the Ocean Biogeographic Information System (OBIS) sponsored by UNESCO's Intergovernmental Oceanographic Commission (IOC); and 5) Understand the linkages between marine biodiversity, ecosystem processes, and the social-economic context of a region. Pilot projects have been implemented within three NOAA National Marine Sanctuaries (Florida Keys, Monterey Bay, and Channel Islands), the wider Santa Barbara Channel, in the Chukchi Sea, and through the Smithsonian's Tennenbaum Marine Observatories Network (TMON) at several sites in the U.S. and collaborating countries. Together, these MBON sites encompass a wide range of marine environments, including deep sea, continental shelves, and coastal habitats including estuaries, wetlands, and coral reefs. The present MBON partners are open to growth of the MBON through additional collaborations. Given these initiatives, GEO BON is proposing an MBON effort that spans from pole to pole, with a pathfinder effort among countries in the Americas. By specializing in coastal ecosystems—where marine biodiversity and people are concentrated and interact most—the MBON and TMON initiatives aim to provide policymakers with the science to

Bioconcentration of TNT and RDX in coastal marine biota.

Science.gov (United States)

Ballentine, Mark; Tobias, Craig; Vlahos, Penny; Smith, Richard; Cooper, Christopher

2015-05-01

The bioconcentration factor (BCF) was measured for 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in seven different marine species of varying trophic levels. Time series and concentration gradient treatments were used for water column and tissue concentrations of TNT, RDX, and their environmentally important derivatives 2-amino-4,6-dintrotoluene (2-ADNT) and 4-amino-2,6-dinitrotoluene (4-ADNT). BCF values ranged from 0.0031 to 484.5 mL g(-1) for TNT and 0.023 to 54.83 mL g(-1) for RDX. The use of log K ow value as an indicator was evaluated by adding marine data from this study to previously published data. For the munitions in this study, log K ow value was a good indicator in the marine environment. The initial uptake and elimination rates of TNT and RDX for Fucus vesiculosus were 1.79 and 0.24 h(-1) for TNT and 0.50 and 0.0035 h(-1) for RDX respectively. Biotransformation was observed in all biota for both TNT and RDX. Biotransformation of TNT favored 4-ADNT over 2-ADNT at ratios of 2:1 for F. vesiculosus and 3:1 for Mytilus edulis. Although RDX derivatives were measureable, the ratios of RDX derivatives were variable with no detectable trend. Previous approaches for measuring BCF in freshwater systems compare favorably with these experiments with marine biota, yet significant gaps on the ultimate fate of munitions within the biota exist that may be overcome with the use stable isotope-labeled munitions substrates.

Marine debris in a World Heritage Listed Brazilian estuary.

Science.gov (United States)

Possatto, Fernanda E; Spach, Henry L; Cattani, André P; Lamour, Marcelo R; Santos, Lilyane O; Cordeiro, Nathalie M A; Broadhurst, Matt K

2015-02-28

Using monthly otter-trawl deployments, spatial and temporal variability among the relative densities of marine debris were assessed in the Paranaguá estuarine complex; a subtropical World Heritage Listed area in southern Brazil. During 432 deployments over 12 months, 291 marine debris items were identified; of which most (92%) were plastic, and more specifically shopping bags, food packages, candy wrappers and cups typically >21 mm long. The most contaminated sectors were those closest to Paranaguá city and the adjacent port, and had up to 23.37±3.22 pieces ha(-1). Less urbanized sectors had between 12.84±1.49 and 9.32±1.10 pieces ha(-1). Contamination did not vary between dry or wet seasons, but rather was probably affected by consistent urban disposal and localized hydrological processes. Marine debris might be minimized by using more environment friendly materials, however a concrete solution requires adequately integrating local government and civil society. Copyright © 2014 Elsevier Ltd. All rights reserved.

Enzymatic Processes in Marine Biotechnology.

Science.gov (United States)

Trincone, Antonio

2017-03-25

In previous review articles the attention of the biocatalytically oriented scientific community towards the marine environment as a source of biocatalysts focused on the habitat-related properties of marine enzymes. Updates have already appeared in the literature, including marine examples of oxidoreductases, hydrolases, transferases, isomerases, ligases, and lyases ready for food and pharmaceutical applications. Here a new approach for searching the literature and presenting a more refined analysis is adopted with respect to previous surveys, centering the attention on the enzymatic process rather than on a single novel activity. Fields of applications are easily individuated: (i) the biorefinery value-chain, where the provision of biomass is one of the most important aspects, with aquaculture as the prominent sector; (ii) the food industry, where the interest in the marine domain is similarly developed to deal with the enzymatic procedures adopted in food manipulation; (iii) the selective and easy extraction/modification of structurally complex marine molecules, where enzymatic treatments are a recognized tool to improve efficiency and selectivity; and (iv) marine biomarkers and derived applications (bioremediation) in pollution monitoring are also included in that these studies could be of high significance for the appreciation of marine bioprocesses.

75 FR 72655 - Marine Sanitation Device Discharge Regulations for the Florida Keys National Marine Sanctuary

Science.gov (United States)

2010-11-26

... National Marine Sanctuary AGENCY: Office of National Marine Sanctuaries (ONMS), National Oceanic and... the regulations for the Florida Keys National Marine Sanctuary (FKNMS or sanctuary) by eliminating the exemption that allows discharges from within the boundary of the sanctuary of biodegradable effluent...

Present status of marine environmental radioactivity survey in the sea of Japan

International Nuclear Information System (INIS)

Matsuoka, H.

1994-01-01

Science and Technology Agency has been conducting some Marine Environmental Radioactivity Surveys around Japan in cooperation with the relevant organizations (Maritime Safety Agency, Japan Meteorological Agency, Fishery Agency, National Institute of Radiological Sciences, Japan Marine Science and Technology Center, Japan Chemical Analysis Center and Marine Ecology Research Institute). Several artificial radionuclides have been detected but the main origin is supposed to be fall-out. The level trend of marine environmental radioactivity has no anomalies excepting the effect of Chernobyl Accident. The data summarized here are as follows. 1. Marine Environmental Survey of Fisheries near the Nuclear Power Stations, 2. Past Data of Marine Environmental Radioactivity around Japan (Apr. 1982 - Mar. 1991), 3. Marine Environmental Survey of the Sea of Japan (spring, 1993), 4. Marine Environmental Survey of the Sea of Japan (autumn, 1993). In addition, JAPAN-KOREA-RUSSIA JOINT EXPEDITION in the Sea of Japan will start in the middle of March. We are expecting to get valuable data through the EXPEDITION. (J.P.N.)

Prevalence of marine debris in marine birds from the North Atlantic.

Science.gov (United States)

Provencher, Jennifer F; Bond, Alexander L; Hedd, April; Montevecchi, William A; Muzaffar, Sabir Bin; Courchesne, Sarah J; Gilchrist, H Grant; Jamieson, Sarah E; Merkel, Flemming R; Falk, Knud; Durinck, Jan; Mallory, Mark L

2014-07-15

Marine birds have been found to ingest plastic debris in many of the world's oceans. Plastic accumulation data from necropsies findings and regurgitation studies are presented on 13 species of marine birds in the North Atlantic, from Georgia, USA to Nunavut, Canada and east to southwest Greenland and the Norwegian Sea. Of the species examined, the two surface plungers (great shearwaters Puffinus gravis; northern fulmars Fulmarus glacialis) had the highest prevalence of ingested plastic (71% and 51%, respectively). Great shearwaters also had the most pieces of plastics in their stomachs, with some individuals containing as many of 36 items. Seven species contained no evidence of plastic debris. Reporting of baseline data as done here is needed to ensure that data are available for marine birds over time and space scales in which we see changes in historical debris patterns in marine environments (i.e. decades) and among oceanographic regions. Copyright © 2014 Elsevier Ltd. All rights reserved.

Transposable elements are enriched within or in close proximity to xenobiotic-metabolizing cytochrome P450 genes

Directory of Open Access Journals (Sweden)

Li Xianchun

2007-03-01

Full Text Available Abstract Background Transposons, i.e. transposable elements (TEs, are the major internal spontaneous mutation agents for the variability of eukaryotic genomes. To address the general issue of whether transposons mediate genomic changes in environment-adaptation genes, we scanned two alleles per each of the six xenobiotic-metabolizing Helicoverpa zea cytochrome P450 loci, including CYP6B8, CYP6B27, CYP321A1, CYP321A2, CYP9A12v3 and CYP9A14, for the presence of transposon insertions by genome walking and sequence analysis. We also scanned thirteen Drosophila melanogaster P450s genes for TE insertions by in silico mapping and literature search. Results Twelve novel transposons, including LINEs (long interspersed nuclear elements, SINEs (short interspersed nuclear elements, MITEs (miniature inverted-repeat transposable elements, one full-length transib-like transposon, and one full-length Tcl-like DNA transpson, are identified from the alleles of the six H. zea P450 genes. The twelve transposons are inserted into the 5'flanking region, 3'flanking region, exon, or intron of the six environment-adaptation P450 genes. In D. melanogaster, seven out of the eight Drosophila P450s (CYP4E2, CYP6A2, CYP6A8, CYP6A9, CYP6G1, CYP6W1, CYP12A4, CYP12D1 implicated in insecticide resistance are associated with a variety of transposons. By contrast, all the five Drosophila P450s (CYP302A1, CYP306A1, CYP307A1, CYP314A1 and CYP315A1 involved in ecdysone biosynthesis and developmental regulation are free of TE insertions. Conclusion These results indicate that TEs are selectively retained within or in close proximity to xenobiotic-metabolizing P450 genes.