Development of genomic array footprinting for identification of conditionally essential genes in Streptococcus pneumoniae.

ABSTRACT: Streptococcus pneumoniae is a major cause of serious infections such as pneumonia and meningitis in both children and adults worldwide. Here, we describe the development of a high-throughput, genome-wide technique, genomic array footprinting (GAF), for the identification of genes essential for this bacterium at various stages during infection. GAF enables negative screens by means of a combination of transposon mutagenesis and microarray technology for the detection of transposon insertion sites. We tested several methods for the identification of transposon insertion sites and found that amplification of DNA adjacent to the insertion site by PCR resulted in nonreproducible results, even when combined with an adapter. However, restriction of genomic DNA followed directly by in vitro transc

SUBMITTER: Bijlsma JJ 

PROVIDER: S-EPMC1828782 | biostudies-literature | 2007 Mar

REPOSITORIES: biostudies-literature