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PMID:2687254
| Citation |
Janssen, DB, Pries, F, van der Ploeg, J, Kazemier, B, Terpstra, P and Witholt, B (1989) Cloning of 1,2-dichloroethane degradation genes of Xanthobacter autotrophicus GJ10 and expression and sequencing of the dhlA gene. J. Bacteriol. 171:6791-9 |
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| 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. |
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| Keywords |
Amino Acid Sequence; Base Sequence; Cloning, Molecular/methods; Escherichia coli/genetics; Ethylene Dichlorides/metabolism; Gene Expression; Genes, Bacterial; Gram-Negative Aerobic Bacteria/enzymology; Gram-Negative Aerobic Bacteria/genetics; Hydrocarbons, Chlorinated/metabolism; Hydrolases/genetics; Molecular Sequence Data; Plasmids; Pseudomonas/genetics; Restriction Mapping; Sequence Homology, Nucleic Acid; Species Specificity; Xanthomonas/genetics |
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Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0018786: haloalkane dehalogenase activity |
ECO:0000316: |
F |
Fig. 3: It was found that DhlA is highly expressed in strains lacking dehalogenase activity and strains with impaired dicholoethane utilization abilities. |
Missing: with/from | ||||
See also
References
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