Molecular Cloning and Functional Characterization of a RecA Analog from Pseudomonas Stutzeri and Construction of a P. Stutzeri RecA Mutant

Overview

Journal Antonie Van Leeuwenhoek

Publisher Springer

Specialty Microbiology

Date 1991 Feb 1

PMID 1854185

Citations 7

Authors

B Vosman

K J Hellingwerf

Affiliations

Soon will be listed here.

Abstract

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 PvuII-BglII 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.

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References

Nilsson B, Uhlen M, Josephson S, Gatenbeck S, Philipson L . An improved positive selection plasmid vector constructed by oligonucleotide mediated mutagenesis. Nucleic Acids Res. 1983; 11(22):8019-30. PMC: 326556. DOI: 10.1093/nar/11.22.8019. View

de Vos W, de Vries S, Venema G . Cloning and expression of the Escherichia coli recA gene in Bacillus subtilis. Gene. 1983; 25(2-3):301-8. DOI: 10.1016/0378-1119(83)90234-2. View

Lovett Jr C, Love P, Yasbin R . Competence-specific induction of the Bacillus subtilis RecA protein analog: evidence for dual regulation of a recombination protein. J Bacteriol. 1989; 171(5):2318-22. PMC: 209904. DOI: 10.1128/jb.171.5.2318-2322.1989. View

Ish-Horowicz D, Burke J . Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981; 9(13):2989-98. PMC: 327326. DOI: 10.1093/nar/9.13.2989. View

Barcak G, Tomb J, Laufer C, Smith H . Two Haemophilus influenzae Rd genes that complement the recA-like mutation rec-1. J Bacteriol. 1989; 171(5):2451-7. PMC: 209920. DOI: 10.1128/jb.171.5.2451-2457.1989. View

JUNI E . Interspecies transformation of Acinetobacter: genetic evidence for a ubiquitous genus. J Bacteriol. 1972; 112(2):917-31. PMC: 251504. DOI: 10.1128/jb.112.2.917-931.1972. View

Little J, Mount D . The SOS regulatory system of Escherichia coli. Cell. 1982; 29(1):11-22. DOI: 10.1016/0092-8674(82)90085-x. View

Clark A . Recombination deficient mutants of E. coli and other bacteria. Annu Rev Genet. 1973; 7:67-86. DOI: 10.1146/annurev.ge.07.120173.000435. View

Boyer H . A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969; 41(3):459-72. DOI: 10.1016/0022-2836(69)90288-5. View

10.

Kooistra J, Setlow J . Similarity in properties and mapping of three Rec mutants of Haemophilus influenzae. J Bacteriol. 1976; 127(1):327-33. PMC: 233065. DOI: 10.1128/jb.127.1.327-333.1976. View

11.

Better M, Helinski D . Isolation and characterization of the recA gene of Rhizobium meliloti. J Bacteriol. 1983; 155(1):311-6. PMC: 217682. DOI: 10.1128/jb.155.1.311-316.1983. View

12.

Vieira J, Messing J . The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982; 19(3):259-68. DOI: 10.1016/0378-1119(82)90015-4. View

13.

Stewart G, Carlson C . The biology of natural transformation. Annu Rev Microbiol. 1986; 40:211-35. DOI: 10.1146/annurev.mi.40.100186.001235. View

14.

Love P, Yasbin R . Genetic characterization of the inducible SOS-like system of Bacillus subtilis. J Bacteriol. 1984; 160(3):910-20. PMC: 215796. DOI: 10.1128/jb.160.3.910-920.1984. View

15.

Walker G . Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev. 1984; 48(1):60-93. PMC: 373003. DOI: 10.1128/mr.48.1.60-93.1984. View

16.

Love P, Lyle M, Yasbin R . DNA-damage-inducible (din) loci are transcriptionally activated in competent Bacillus subtilis. Proc Natl Acad Sci U S A. 1985; 82(18):6201-5. PMC: 391020. DOI: 10.1073/pnas.82.18.6201. View

17.

Bolivar F, Rodriguez R, Greene P, Betlach M, Heyneker H, Boyer H . Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977; 2(2):95-113. View

18.

Resnick D, Nelson D . Cloning and characterization of the Aeromonas caviae recA gene and construction of an A. caviae recA mutant. J Bacteriol. 1988; 170(1):48-55. PMC: 210604. DOI: 10.1128/jb.170.1.48-55.1988. View

19.

Berson A, Hudson D, Waleh N . Cloning and characterization of the recA gene of Aquaspirillum magnetotacticum. Arch Microbiol. 1989; 152(6):567-71. DOI: 10.1007/BF00425488. View

20.

Vieira J, Messing J . Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985; 33(1):103-19. DOI: 10.1016/0378-1119(85)90120-9. View