|Ricke, Steven - TEXAS A&M UNIVERSITY|
Submitted to: American Society of Microbiologists Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: May 22, 2001
Publication Date: N/A
Technical Abstract: An increasing number of microbial genomes have been completely sequenced. However, the biological functions of the sequences are largely unknown for the microorganisms, mainly due to the lack of a comprehensive method for functional analysis of microbial genomes. We devised a quantitative method for functional analysis of microbial genomes in genome-wide scale using transposon mutagenesis and sequence tags that are defined by their positions in the transposon-flanking sequences. We demonstrated that the identity and frequency of the sequence tags provide information on the level of each mutant in the pool. Therefore, transposon mutants with competitive disadvantages can be identified by comparing the sequence tag profiles of the mutant pools before and after selection. We applied this method to identify the Tn5 mutants of Salmonella enteritidis that are attenuated during infection in 5-day-old chicks. We identified potential virulence genes that include previously known virulence genes. This method is a powerful approach for categorizing gene function that should be applicable to a variety of microorganisms.