Document Detail


Development of metagenomic DNA shuffling for the construction of a xenobiotic gene.
MedLine Citation:
PMID:  16690231     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Hasna Boubakri; Mélanie Beuf; Pascal Simonet; Timothy M Vogel
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-04-01
Journal Detail:
Title:  Gene     Volume:  375     ISSN:  0378-1119     ISO Abbreviation:  Gene     Publication Date:  2006 Jun 
Date Detail:
Created Date:  2006-06-09     Completed Date:  2006-08-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7706761     Medline TA:  Gene     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  87-94     Citation Subset:  IM    
Affiliation:
Ecologie Microbienne, UMR CNRS 5557, Université Claude Bernard Lyon I, 16 rue Dubois, F-69622 Villeurbanne Cedex, France.
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MeSH Terms
Descriptor/Qualifier:
Bacteria / genetics
Base Sequence
DNA Primers
DNA, Bacterial / genetics*
Genome, Bacterial*
Mutation
Open Reading Frames
Polymerase Chain Reaction
Recombination, Genetic
Xenobiotics / metabolism*
Chemical
Reg. No./Substance:
0/DNA Primers; 0/DNA, Bacterial; 0/Xenobiotics

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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