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      Characterisation of the effect of a simulated hydrocarbon spill on diazotrophs in mangrove sediment mesocosm

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          Abstract

          An analysis of the effect of an oil spill on mangrove sediments was carried out by contamination of mesocosms derived from two different mangroves, one with a history of contamination and one pristine. The association between N 2 fixers and hydrocarbon degradation was assessed using quantitative PCR (qPCR) for the genes rrs and nifH, nifH clone library sequencing and total petroleum hydrocarbon (TPH) quantification using gas chromatography. TPH showed that the microbial communities of both mangroves were able to degrade the hydrocarbons added; however, whereas the majority of oil added to the mesocosm derived from the polluted mangrove was degraded in the 75 days of the experiment, there was only partially degradation in the mesocosm derived from the pristine mangrove. qPCR showed that the addition of oil led to an increase in rrs gene copy numbers in both mesocosms, having almost no effect on the nifH copy numbers in the pristine mangrove. Sequencing of nifH clones indicated that the changes promoted by the oil in the polluted mangrove were greater than those observed in the pristine mesocosm. The main effect observed in the polluted mesocosm was the selection of a single phylotype which is probably adapted to the presence of petroleum. These results, together with previous reports, give hints about the relationship between N 2 fixation and hydrocarbon degradation in natural ecosystems.

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          Molecular Cloning : A Laboratory Manual

          <p>The first two editions of this manual have been mainstays of molecular biology for nearly twenty years, with an unrivalled reputation for reliability, accuracy, and clarity.<br>In this new edition, authors Joseph Sambrook and David Russell have completely updated the book, revising every protocol and adding a mass of new material, to broaden its scope and maintain its unbeatable value for studies in genetics, molecular cell biology, developmental biology, microbiology, neuroscience, and immunology.<br>Handsomely redesigned and presented in new bindings of proven durability, this three–volume work is essential for everyone using today’s biomolecular techniques.<br>The opening chapters describe essential techniques, some well–established, some new, that are used every day in the best laboratories for isolating, analyzing and cloning DNA molecules, both large and small.<br>These are followed by chapters on cDNA cloning and exon trapping, amplification of DNA, generation and use of nucleic acid probes, mutagenesis, and DNA sequencing.<br>The concluding chapters deal with methods to screen expression libraries, express cloned genes in both prokaryotes and eukaryotic cells, analyze transcripts and proteins, and detect protein–protein interactions.<br>The Appendix is a compendium of reagents, vectors, media, technical suppliers, kits, electronic resources and other essential information.<br>As in earlier editions, this is the only manual that explains how to achieve success in cloning and provides a wealth of information about why techniques work, how they were first developed, and how they have evolved. </p>
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            Le piégeage lumineux, moyen d'approche de la faune entomologique d'un grand fleuve (Ephéméroptères, en particulier)

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              Nitrogenase gene diversity and microbial community structure: a cross-system comparison.

              Biological nitrogen fixation is an important source of fixed nitrogen for the biosphere. Microorganisms catalyse biological nitrogen fixation with the enzyme nitrogenase, which has been highly conserved through evolution. Cloning and sequencing of one of the nitrogenase structural genes, nifH, has provided a large, rapidly expanding database of sequences from diverse terrestrial and aquatic environments. Comparison of nifH phylogenies to ribosomal RNA phylogenies from cultivated microorganisms shows little conclusive evidence of lateral gene transfer. Sequence diversity far outstrips representation by cultivated representatives. The phylogeny of nitrogenase includes branches that represent phylotypic groupings based on ribosomal RNA phylogeny, but also includes paralogous clades including the alternative, non-molybdenum, non-vanadium containing nitrogenases. Only a few alternative or archaeal nitrogenase sequences have as yet been obtained from the environment. Extensive analysis of the distribution of nifH phylotypes among habitats indicates that there are characteristic patterns of nitrogen fixing microorganisms in termite guts, sediment and soil environments, estuaries and salt marshes, and oligotrophic oceans. The distribution of nitrogen-fixing microorganisms, although not entirely dictated by the nitrogen availability in the environment, is non-random and can be predicted on the basis of habitat characteristics. The ability to assay for gene expression and investigate genome arrangements provides the promise of new tools for interrogating natural populations of diazotrophs. The broad analysis of nitrogenase genes provides a basis for developing molecular assays and bioinformatics approaches for the study of nitrogen fixation in the environment.
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                Author and article information

                Contributors
                rgtaketani@cena.usp.br
                Journal
                Antonie Van Leeuwenhoek
                Antonie Van Leeuwenhoek
                Springer Netherlands (Dordrecht )
                0003-6072
                1572-9699
                26 May 2009
                October 2009
                : 96
                : 3
                : 343-354
                Affiliations
                [1 ]Laboratório de Ecologia Microbiana Molecular, Instituto de Microbiologia, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ Brazil
                [2 ]Microbial Ecology Department, Center for Ecological and Evolutionary Studies, University of Groningen, Groningen, The Netherlands
                [3 ]Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, 303 Centenário Av., Piracicaba, SP 13400-970 Brazil
                Article
                9351
                10.1007/s10482-009-9351-6
                2729449
                19468855
                3b450743-ee82-46fa-bd4e-641e553d483f
                © The Author(s) 2009
                History
                : 1 April 2009
                : 12 May 2009
                Categories
                Original Paper
                Custom metadata
                © Springer Science+Business Media B.V. 2009

                Microbiology & Virology
                microbial community,mangrove sediment,oil degradation,diazotrophs
                Microbiology & Virology
                microbial community, mangrove sediment, oil degradation, diazotrophs

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