Transposon-mediated random gene disruption with moderate halophilic bacteria and its application for halophilic bacterial siderophore analysis.
Analytical conditions using chromo azurol S was validated for quantification of siderophore in aqueous samples, followed by the characterization of siderophore derived from newly isolated moderately halophilic bacteria. Conditions with good linearity between the absorbance and the siderophore concentration were obtained at a siderophore concentration less than 20 µM, in the wavelength range between 630 and 660 nm with developing time for at least 2 h. Of the halophilic bacteria isolated from Tunisian soil, Halomonas sp., namely strain 21a was selected as siderophore producing halophiles. The strain produced siderophore significantly in the absence of iron in minimal medium. Siderophore-deficient mutant, namely IIa10, of the strain 21a was obtained from gene disruptant library constructed using transposon complex by electroporation. Genomic sequence analysis of the mutant IIa10 revealed that the transposon-inserted gene was TonB-dependent receptor.
School of Bioscience and Biotechnology, Tokyo University of Technology, 1404 Katakura, Hachioji, Tokyo, 192-0982, Japan.
Toru
T
Matsui
School of Bioscience and Biotechnology, Tokyo University of Technology, 1404 Katakura, Hachioji, Tokyo, 192-0982, Japan.
Tomohiko
T
Nishino
D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D016680:Genome, Bacterial; D020632:Halomonas / Q000235:genetics* / Q000378:metabolism; D007501:Iron / Q000378:metabolism; D016254:Mutagenesis, Insertional; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D017262:Siderophores / Q000032:analysis* / Q000096:biosynthesis; D012988:Soil Microbiology
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D017262:Siderophores; D007501:Iron
Halomonas sp.; Halophiles; Siderophore; TonB-dependent receptor; Transposon