Developing a Bacteroides System for Function-Based Screening of DNA from the Human Gut Microbiome.

ABSTRACT: Functional metagenomics is a powerful method that allows the isolation of genes whose role may not have been predicted from DNA sequence. In this approach, first, environmental DNA is cloned to generate metagenomic libraries that are maintained in Escherichia coli, and second, the cloned DNA is screened for activities of interest. Typically, functional screens are carried out using E. coli as a surrogate host, although there likely exist barriers to gene expression, such as lack of recognition of native promoters. Here, we describe efforts to develop Bacteroides thetaiotaomicron as a surrogate host for screening metagenomic DNA from the human gut. We construct a B. thetaiotaomicron-compatible fosmid cloning vector, generate a fosmid clone library using DNA from the human gut, and show succ

SUBMITTER: Lam KN 

PROVIDER: S-EPMC5872301 | biostudies-literature | 2018 May-Jun

REPOSITORIES: biostudies-literature