Development and use of a selectable, broad-host-range reporter transposon for identifying environmentally regulated promoters in bacteria
Year: 2008
Type: article
Abstract: This report describes the development and use of TnKnXSp, a selectable broad-host-range reporter transposon with a promoterless aphA gene. Insertion of TnKnXSp into the chromosome of a kanamycin-susceptible bacterium confers resistance to kanamycin only if aphA is transcribed from an active promoter adjacent to the insertion site. We designed TnKnXSp as a tool for identifying environmentally regulated promoters in bacteria and developed general methods for initial characterization of any TnKnXSp integrant. To identify putative iron-regulated promoters in Corynebacterium diphtheriae, we constructed TnKnXSp integrants and identified a subgroup that expressed kanamycin resistance under low-iron, but not high-iron, conditions. We characterized representative integrants with this phenotype, located the TnKnXSp insertion in each, and demonstrated that transcription of aphA was repressed under high-iron vs. low-iron growth conditions. We also demonstrated that TnKnXSp can be used in bacteria other than C. diphtheriae, including Escherichia coli and Bacillus subtilis. Our findings validate TnKnXSp as a useful tool for identifying environmentally regulated promoters in bacteria.
Source: FEMS Microbiology Letters
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: green
Awards: G1024577694, Control of Bacterial Toxins by Viruses and Plasmids, G1869106726, CONTROL OF BACTERIAL TOXINS BY VIRUSES AND PLASMIDS, G1946161438, Molecular Pathogenesis of Infectious Diseases, G783754770, NIAMS-Research Training in Dermatology, University of Colorado Cancer Center Support Grant
OpenAlex ID: W2073743176