Preparation of fosmid libraries and functional metagenomic analysis of microbial community DNA.
One of the most important challenges in contemporary microbial ecology is to assign a functional role to the large number of novel genes discovered through large-scale sequencing of natural microbial communities that lack similarity to genes of known function. Functional screening of metagenomic libraries, that is, screening environmental DNA clones for the ability to confer an activity of interest to a heterologous bacterial host, is a promising approach for bridging the gap between metagenomic DNA sequencing and functional characterization. Here, we describe methods for isolating environmental DNA and constructing metagenomic fosmid libraries, as well as methods for designing and implementing successful functional screens of such libraries.
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. Electronic address: chon@mit.edu.
Asunción
A
Martínez
Marcia S
MS
Osburne
D004269:DNA, Bacterial; D005822:Genetic Vectors; D015698:Genomic Library; D056186:Metagenomics / Q000379:methods*; D059013:Microbial Consortia / Q000235:genetics*
D004269:DNA, Bacterial
DNA library; Environmental; Enzymatic assay; Fosmid; Functional screens; High-throughput assay; Metagenomic