A genome-wide inducible phenotypic screen identifies antisense RNA constructs silencing Escherichia coli essential genes.

ABSTRACT: Regulated antisense RNA (asRNA) expression has been employed successfully in Gram-positive bacteria for genome-wide essential gene identification and drug target determination. However, there have been no published reports describing the application of asRNA gene silencing for comprehensive analyses of essential genes in Gram-negative bacteria. In this study, we report the first genome-wide identification of asRNA constructs for essential genes in Escherichia coli. We screened 250 000 library transformants for conditional growth inhibitory recombinant clones from two shotgun genomic libraries of E. coli using a paired-termini expression vector (pHN678). After sequencing plasmid inserts of 675 confirmed inducer sensitive cell clones, we identified 152 separate asRNA constructs of which 134

SUBMITTER: Meng J 

PROVIDER: S-EPMC3855647 | biostudies-literature | 2012 Apr

REPOSITORIES: biostudies-literature