GONUTS has been updated to MW1.31 Most things seem to be working but be sure to report problems.
PMID:3036764
| Citation |
Streber, WR, Timmis, KN and Zenk, MH (1987) Analysis, cloning, and high-level expression of 2,4-dichlorophenoxyacetate monooxygenase gene tfdA of Alcaligenes eutrophus JMP134. J. Bacteriol. 169:2950-5 |
|---|---|
| Abstract |
Plasmid pJP4 of Alcaligenes eutrophus JMP134 contains all genes for the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D). Five of these genes, tfdB, tfdC, tfdD, tfdE, and tfdF, have recently been localized and cloned (R. H. Don, A. J. Weightman, H.-J. Knackmuss, and K. N. Timmis, J. Bacteriol. 161:85-90, 1985). Gene tfdA, which codes for the 2,4-D monooxygenase, has now been found by mutagenesis with transposon Tn5. A 3-kilobase fragment of pJP4 cloned in a broad-host-range vector could complement the 2,4-D-negative phenotype of two mutants which lacked 2,4-D monooxygenase activity. The cloned tfdA gene was also transferred to A. eutrophus JMP222, which is a cured derivative of JMP134. The recombinant strain could utilize phenoxyacetic acid as a sole source of carbon and energy. Pseudomonas sp. strain B13, containing the cloned tfdA, was able to degrade phenoxyacetic acid and 4-chlorophenoxyacetic acid. Gene tfdA was subcloned and analyzed by deletions. Expression of 2,4-D monooxygenase in Escherichia coli containing a 1.4-kilobase subfragment was demonstrated by radioisotopic enzyme assay, and a protein of 32,000-dalton molecular mass was detected by labeling experiments. A 2-kilobase subfragment containing tfdA has been sequenced. Sequence analysis revealed an open reading frame of 861 bases which was identified as the coding region of tfdA by insertion mutagenesis. |
| Links | |
| Keywords |
Alcaligenes/genetics; Base Sequence; Chromosome Deletion; Cloning, Molecular; DNA Transposable Elements; DNA, Bacterial/genetics; Gene Expression Regulation; Genes, Bacterial; Mutation; Oxygenases/genetics; Plasmids; Pseudomonas/genetics |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
|
Contributes to |
GO:0018602: 2,4-dichlorophenoxyacetate alpha-ketoglutarate dioxygenase activity |
ECO:0000314: |
F |
TfdA codes for the 2,4-D monooxygenase. Figure 4 shows omega mutagenesis of the tfdA gene. An omega fragment containing stop signals was inserted into the coding region of a tfdA plasmid. The results were a truncated tfdA product for the plasmids which the omega fragment was inserted, in turn disrupting the enzymatic activity for the 2,4-D monooxygenase. |
complete | |||
See also
References
See Help:References for how to manage references in GONUTS.
