Journal
NATURE BIOTECHNOLOGY
Volume 18, Issue 7, Pages 740-745Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/77305
Keywords
essential genes; functional genomics; transposon mutagenesis; Vibrio cholerae; cholera
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Funding
- NIAID NIH HHS [AI-26289] Funding Source: Medline
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Identification of genes that encode essential products provides a promising approach to validation of new antibacterial drug targets. We have developed a mariner-based transposon, TnAraOut, that allows efficient identification and characterization of essential genes by transcriptionally fusing them to an outward-facing, arabinose-inducible promoter, P-BAD, located at one end of the transposon. In the absence of arabinose, such TnAraOut fusion strains display pronounced growth defects. Of a total of 16 arabinose-dependent TnAraOut mutants characterized in Vibrio cholerae, four were found to carry insertions upstream of known essential genes (gyrB, proRS, ileRS, and aspRS) whereas the other strains carried insertions upstream of known and hypothetical genes not previously shown to encode essential gene products. One of the essential genes identified by this analysis appears to be unique to V. cholerae and thus may represent an example of a species-specific drug target.
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