Tn5 Tagging of the Phenol-degrading Gene on the Chromosome of Pseudomonas putida
- Authors
- Han E.-M.; Jung Y.-H.; On H.-Y.; Lee M.-S.; Min K.-H.; Yang Y.-K.; Kim Y.; Kim C.-K.; Lee K.-S.
- Issue Date
- Feb-1997
- Publisher
- Springer Verlag
- Citation
- Molecules and Cells, v.7, no.1, pp 40 - 44
- Pages
- 5
- Journal Title
- Molecules and Cells
- Volume
- 7
- Number
- 1
- Start Page
- 40
- End Page
- 44
- URI
- https://scholarworks.sookmyung.ac.kr/handle/2020.sw.sookmyung/150525
- ISSN
- 1016-8478
0219-1032
- Abstract
- Transposon mutagenesis was performed by the method of conjugational transfer in order to identify and characterize genes encoding enzymes involved in the pathway of phenol utilization as a carbon source. Escherichia coli, which carries the Tn5-132, Was mated with Pseudomonas putida SM25 as a host. We selected a mutant that could not utilize phenol as a carbon source. Chromosomal integration of the transposon was confirmed by Southern analysis, successfully tagging the gene related to a phenol-utilizing pathway. By cell-free enzyme and genetic complementation assays, the inactivated enzyme through the mutation of the corresponding gene was identified as the catB gene, which encodes a cis,cis-muconate lactonizing enzyme.
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