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PMID: 9251187 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Use of a promoterless lacZ gene insertion to investigate chitinase gene expression in the marine bacterium Pseudoalteromonas sp. strain S9.

Applied and environmental microbiology ·Vol. 63 ·No. 8 ·1997-08-00 ·Pages 2989-96

Techkarnjanaruk S, Pongpattanakitshote S, Goodman AE

Abstract

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.

MeSH Terms
Acetylglucosamine/metabolism Amino Acid Sequence Base Sequence Cadmium/metabolism Carbon Dioxide/metabolism Chitin/metabolism Chitinases/genetics,metabolism Cloning, Molecular DNA Transposable Elements Gene Expression Regulation, Bacterial Gene Expression Regulation, Enzymologic Genes, Reporter Glucose/metabolism Gram-Negative Aerobic Bacteria/classification,genetics Heat-Shock Response Lac Operon Molecular Sequence Data Phylogeny Promoter Regions, Genetic RNA, Ribosomal, 16S/genetics Repressor Proteins Sequence Alignment Sequence Analysis, DNA Sequence Homology, Amino Acid Ultraviolet Rays beta-Galactosidase/metabolism
Chemicals
DNA Transposable Elements RNA, Ribosomal, 16S Repressor Proteins Cadmium Chitin Carbon Dioxide Chitinases beta-Galactosidase Glucose Acetylglucosamine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Techkarnjanaruk S
School of Biological Sciences, Flinders University of South Australia, Adelaide, Australia. S.Techkarn@flinders.edu.au
Pongpattanakitshote S
Goodman A E
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/aem.63.8.2989-2996.1997
Email
s.techkarn@flinders.edu.au
Published
1997-08-00
Pages
2989-96
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC168598
Subset
IM
Databases
GENBANK
U80834
External Links
PubMed source DOI: 10.1128/aem.63.8.2989-2996.1997 PMC Full Text View journal details
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