Heterologous viral expression systems in fosmid vectors increase the functional analysis potential of metagenomic libraries.

ABSTRACT: The extraordinary potential of metagenomic functional analyses to identify activities of interest present in uncultured microorganisms has been limited by reduced gene expression in surrogate hosts. We have developed vectors and specialized E. coli strains as improved metagenomic DNA heterologous expression systems, taking advantage of viral components that prevent transcription termination at metagenomic terminators. One of the systems uses the phage T7 RNA-polymerase to drive metagenomic gene expression, while the other approach uses the lambda phage transcription anti-termination protein N to limit transcription termination. A metagenomic library was constructed and functionally screened to identify genes conferring carbenicillin resistance to E. coli. The use of these enhanced expressi

SUBMITTER: Terron-Gonzalez L 

PROVIDER: S-EPMC3551230 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature