Identification of 113 conserved essential genes using a high-throughput gene disruption system in Streptococcus pneumoniae.

ABSTRACT: The recent availability of bacterial genome sequence information permits the identification of conserved genes that are potential targets for novel antibiotic drug discovery. Using a coupled bioinformatic/experimental approach, a list of candidate conserved genes was generated using a Microbial Concordance bioinformatics tool followed by a targeted disruption campaign. Pneumococcal sequence data allowed for the design of precise PCR primers to clone the desired gene target fragments into the pEVP3 'suicide vector'. An insertion-duplication approach was employed that used the pEVP3 constructs and resulted in the introduction of a selectable chloramphenicol resistance marker into the chromosome. In the case of non-essential genes, cells can survive the disruption and form chloramphenicol-res

SUBMITTER: Thanassi JA 

PROVIDER: S-EPMC135739 | biostudies-literature | 2002 Jul

REPOSITORIES: biostudies-literature