Isolation of Citrobacter sp. mutants defective in decolorization of brilliant green by transposon mutagenesis.
To identify genes involved in the decolorization of brilliant green, we isolated random mutants generated by transposon insertion in brilliant green-decolorizing bacterium, Citrobacter sp. The resulting mutant bank yielded 19 mutants with a complete defect in terms of the brilliant green color removing ability. Southern hybridization with a Tn5 fragment as a probe showed a single hybridized band in 7 mutants and these mutants appeared to have insertions at different sites of the chromosome. Tn5-inserted genes were isolated and the DNA sequence flanking Tn5 was determined. By comparing these with a sequence database, putative protein products encoded by bg genes were identified as follows: bg 3 as a LysR-type regulatory protein; bg 11 as a MalG protein in the maltose transport system; bg 14 as an oxidoreductase; and bg 17 as an ABC transporter. The sequences deduced from the three bg genes, bg 2, bg 7 and bg 16, showed no significant similarity to any protein with a known function, suggesting that these three bg genes may encode unidentified proteins responsible for the decolorization of brilliant green.
Jang
Moon-Sun
MS
Faculty of Biotechnology, College of Natural Resources and Life Science, Dong-A University, Busan 604-714, Korea.
Lee
Young-Mi
YM
Choi
Yong-Lark
YL
Cho
Young-Su
YS
Lee
Young-Choon
YC
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D015139:Blotting, Southern; D002954:Citrobacter / Q000235:genetics* / Q000378:metabolism; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D029968:Escherichia coli Proteins / Q000235:genetics; D017353:Gene Deletion; D016254:Mutagenesis, Insertional; D009154:Mutation; D010088:Oxidoreductases / Q000235:genetics; D000644:Quaternary Ammonium Compounds / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014157:Transcription Factors / Q000235:genetics
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C079123:MalG protein, E coli; D000644:Quaternary Ammonium Compounds; D014157:Transcription Factors; C073470:LysR protein, Bacteria; D010088:Oxidoreductases; C006798:brilliant green