Use of a promoterless lacZ gene insertion to investigate chitinase gene expression in the marine bacterium Pseudoalteromonas sp. strain S9.
Sequence data for genes encoding 16S rRNA indicated that the marine strain previously named Pseudomonas sp. strain S9 would be better identified as a Pseudoalteromonas sp. By use of transposon mutagenesis, a chitinase-negative mutant of S9 with a lacZ reporter gene insertion was isolated. Part of the interrupted gene was cloned and sequenced. The deduced amino acid sequence had homology to sequences of bacterial chitinases. Expression of the chitinase gene promoter was quantified by measuring the lacZ reporter gene product, beta-galactosidase, beta-Galactosidase production was induced 10-fold by N-acetylglucosamine and 3-fold by chitin in minimal medium. Repression of beta-galactosidase synthesis was observed in rich medium either with or without chitin but was not observed in minimal medium containing glucose. The chitinase gene promoter was induced by starvation and higher-than-ambient levels of carbon dioxide but not by cadmium ion, heat or cold shock, or UV exposure.
Techkarnjanaruk
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School of Biological Sciences, Flinders University of South Australia, Adelaide, Australia. S.Techkarn@flinders.edu.au
Pongpattanakitshote
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S
Goodman
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D000117:Acetylglucosamine / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D002104:Cadmium / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D017930:Genes, Reporter; D005947:Glucose / Q000378:metabolism; D006088:Gram-Negative Aerobic Bacteria / Q000145:classification / Q000235:genetics*; D018869:Heat-Shock Response; D007763:Lac Operon; D008969:Molecular Sequence Data; D010802:Phylogeny; D011401:Promoter Regions, Genetic; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012097:Repressor Proteins; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014466:Ultraviolet Rays; D001616:beta-Galactosidase / Q000378:metabolism
D004251:DNA Transposable Elements; D012336:RNA, Ribosomal, 16S; D012097:Repressor Proteins; D002104:Cadmium; D002686:Chitin; D002245:Carbon Dioxide; D002688:Chitinases; D001616:beta-Galactosidase; D005947:Glucose; D000117:Acetylglucosamine