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Cloning and expression in Escherichia coli of a recA-like gene from Vibrio cholerae

✍ Scribed by Paul, Kalidas ;Ghosh, Subrata K. ;Das, Jyotirmoy

Publisher: Springer
Year: 1986
Tongue: English
Weight: 645 KB
Volume: 203
Category: Article
ISSN: 0026-8925
DOI: 10.1007/bf00330384

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✦ Synopsis

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 lambda repressor. Induction of a 39,000 dalton protein in UV-irradiated V. cholerae cells was demonstrated.

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