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Gene
Article . 2006 . Peer-reviewed
License: Elsevier TDM
Data sources: Crossref
image/svg+xml Jakob Voss, based on art designer at PLoS, modified by Wikipedia users Nina and Beao Closed Access logo, derived from PLoS Open Access logo. This version with transparent background. http://commons.wikimedia.org/wiki/File:Closed_Access_logo_transparent.svg Jakob Voss, based on art designer at PLoS, modified by Wikipedia users Nina and Beao
HAL INRAE
Article . 2006
Data sources: HAL INRAE
Gene
Article . 2006
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Development of metagenomic DNA shuffling for the construction of a xenobiotic gene

Authors: Boubakri, Hasna; Beuf, M.; Simonet, Pascal; Vogel, Timothy;

Development of metagenomic DNA shuffling for the construction of a xenobiotic gene

Abstract

We describe a metagenomic DNA shuffling process by combining protein engineering process mutation generator and the high potential diversity of metagenomic DNA derived from the environment. Numerous previous shuffling processes attempted to recombine more or less related parental sequences. At the same time, metagenomic approaches unveiled a huge diversity of DNA sequences and genomes, which have not yet been identified to date. In this study, we attempted to combine these two approaches in order to regenerate a novel gene. Here, we present the possibility that DNA fragments from an entire microbial community (metagenome) might be available for the creation of novel genes capable of degrading pollutants. Metagenomic DNA extracted from non-polluted soil was shuffled in vitro to recreate the linA gene responsible for the first steps of lindane degradation. In this work, 74% of the ORF came from separate subsets of the metagenomic pool from a lindane-free and linA-free soil. Our results demonstrate that microbial community genetic diversity can serve as a source for novel gene construction during in vitro manipulation. This in vitro gene construction might also simulate the mosaic nature of novel genes. This demonstration might lead to other attempts to mimic bacterial adaptation and to construct degradative genes for novel compounds not yet released into the environment.

Country
France
Keywords

HALOALKANE DEHALOGENASE, DNA, Bacterial, LIN GENES, DIRECTED EVOLUTION, FOREIGN DNA, Polymerase Chain Reaction, Xenobiotics, Open Reading Frames, GAMMA-HEXACHLOROCYCLOHEXANE, SPHINGOMONAS-PAUCIMOBILIS, DNA Primers, Recombination, Genetic, Bacteria, Base Sequence, MOLECULAR EVOLUTION, GENOME, SOIL, Mutation, BACTERIAL COMMUNITIES, Genome, Bacterial, [SPI.NRJ] Engineering Sciences [physics]/Electric power

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
43
Top 10%
Top 10%
Top 10%
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