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PMID: 2687254 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Cloning of 1,2-dichloroethane degradation genes of Xanthobacter autotrophicus GJ10 and expression and sequencing of the dhlA gene.

Journal of bacteriology ·Vol. 171 ·No. 12 ·1989-12-00 ·Pages 6791-9

Janssen DB, Pries F, van der Ploeg J, Kazemier B, Terpstra P, Witholt B

Abstract

A gene bank from the chlorinated hydrocarbon-degrading bacterium Xanthobacter autotrophicus GJ10 was prepared in the broad-host-range cosmid vector pLAFR1. By using mutants impaired in dichloroethane utilization and strains lacking dehalogenase activities, several genes involved in 1,2-dichloroethane metabolism were isolated. The haloalkane dehalogenase gene dhlA was subcloned, and it was efficiently expressed from its own constitutive promoter in strains of a Pseudomonas sp., Escherichia coli, and a Xanthobacter sp. at levels up to 30% of the total soluble cellular protein. A 3-kilobase-pair BamHI DNA fragment on which the dhlA gene is localized was sequenced. The haloalkane dehalogenase gene was identified by the known N-terminal amino acid sequence of its product and found to encode a 310-amino-acid protein of molecular weight 35,143. Upstream of the dehalogenase gene, a good ribosome-binding site and two consensus E. coli promoter sequences were present.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular/methods Escherichia coli/genetics Ethylene Dichlorides/metabolism Gene Expression Genes, Bacterial Gram-Negative Aerobic Bacteria/enzymology,genetics Hydrocarbons, Chlorinated/metabolism Hydrolases/genetics Molecular Sequence Data Plasmids Pseudomonas/genetics Restriction Mapping Sequence Homology, Nucleic Acid Species Specificity Xanthomonas/genetics
Chemicals
Ethylene Dichlorides Hydrocarbons, Chlorinated ethylene dichloride Hydrolases haloalkane dehalogenase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Janssen D B
Department of Biochemistry, University of Groningen, The Netherlands.
Pries F
van der Ploeg J
Kazemier B
Terpstra P
Witholt B
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.171.12.6791-6799.1989
Published
1989-12-00
Pages
6791-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC210578
Subset
IM
Databases
GENBANK
M26950
External Links
PubMed source DOI: 10.1128/jb.171.12.6791-6799.1989 PMC Full Text View journal details
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