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Characterization of new bacterial catabolic genes and mobile genetic elements by high throughput genetic screening of a soil metagenomic library.

Authors :
Jacquiod, Samuel
Demanèche, Sandrine
Franqueville, Laure
Ausec, Luka
Xu, Zhuofei
Delmont, Tom O.
Dunon, Vincent
Cagnon, Christine
Mandic-Mulec, Ines
Vogel, Timothy M.
Simonet, Pascal
Source :
Journal of Biotechnology. Nov2014, Vol. 190, p18-29. 12p.
Publication Year :
2014

Abstract

A mix of oligonucleotide probes was used to hybridize soil metagenomic DNA from a fosmid clone library spotted on high density membranes. The pooled radio-labeled probes were designed to target genes encoding glycoside hydrolases GH18, dehalogenases, bacterial laccases and mobile genetic elements (integrases from integrons and insertion sequences). Positive hybridizing spots were affiliated to the corresponding clones in the library and the metagenomic inserts were sequenced. After assembly and annotation, new coding DNA sequences related to genes of interest were identified with low protein similarity against the closest hits in databases. This work highlights the sensitivity of DNA/DNA hybridization techniques as an effective and complementary way to recover novel genes from large metagenomic clone libraries. This study also supports that some of the identified catabolic genes might be associated with horizontal transfer events. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
01681656
Volume :
190
Database :
Academic Search Index
Journal :
Journal of Biotechnology
Publication Type :
Academic Journal
Accession number :
99611430
Full Text :
https://doi.org/10.1016/j.jbiotec.2014.03.036