Molecular cloning and functional characterization of arecAanalog fromPseudomonas stutzeriand construction of aP. stutzeri recAmutant
✍ Scribed by Ben Vosman; Klaas J. Hellingwerf
- Publisher
- Springer Netherlands
- Year
- 1991
- Tongue
- English
- Weight
- 765 KB
- Volume
- 59
- Category
- Article
- ISSN
- 0003-6072
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✦ Synopsis
A recombinant plasmid carrying the Pseudomonas stutzeri recA gene was isolated by complementation of the Escherichia coli recA13 mutation. Subcloning experiments showed that the gene was located on a 1500 bp PvulI-BgllI fragment. The cloned gene complements an E. coli recA mutant for resistance to Methylmethanosulphonate (MMS) and UV irradiation. It was also capable of restoring the recombination proficiency in that mutant.
The cloned fragment was used to construct a recA deletion mutant of P. stutzeri. This mutant too was shown to be sensitive towards MMS and UV irradiation. The mutant strain was found to be completely deficient in natural transformation with chromosomal DNA, due to the impairment in homologous recombination.