Cloning and expression in Escherichia coli of a recA-like gene from Vibrio cholerae |
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Authors: | R Lützelschwab G Müller B Wälder O Schmidt R Fürbass and B Mechler |
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Institution: | (1) Institut für Biologie, Universität, Schänzlestraße 1, D-7800 Freiburg, Federal Republic of Germany;(2) Institut für Genetik, Johannes Gutenberg-Universität, Saarstraße 21, Postfach 3980, D-6500 Mainz, Federal Republic of Germany |
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Abstract: | Summary A library containing more than 80% of the Vibrio cholerae genome was constructed by cloning BamH1 restriction fragments into pBR322. Using interspecific complementation of an Escherichia coli recA mutant with plasmids containing the gene bank of V. cholerae, a recA-like gene was identified. The recombinant plasmid, designated as pDP145, contained a 1.45 kb segment of V. cholerae DNA which codes for a protein of molecular weight 39,000. The product of this gene confers methyl methane sulphonate resistance on the E. coli recA mutant, suppresses its ultraviolet (UV) light sensitive phenotype and has proteolytic activity on the phage repressor. Induction of a 39,000 dalton protein in UV-irradiated V. cholerae cells was demonstrated. |
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Keywords: | Genomic library Recombinant plasmid pDP145 recA protein SOS induction UV sensitivity |
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