Open AccessCloning and sequence analysis of a recA ‐like gene from Streptomyces venezuelae ISP5230
Author(s) -
Yao W.,
Vining L.C.
Publication year - 1994
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1994.tb06802.x
Subject(s) - biology , plasmid , genetics , microbiology and biotechnology , molecular cloning , open reading frame , gene , nucleic acid sequence , multiple cloning site , sequence analysis , escherichia coli , cloning (programming) , genomic library , peptide sequence , recombinant dna , expression vector , computer science , programming language
When genomic DNA fragments from Streptomyces venezuelae ISP5230 were probed at moderate stringency with recA from Mycobacterium tuberculosis a 2.0‐kb Sma I fragment was identified. The fragment was isolated by cloning a Bam HI digest of S. venezuelae DNA in pHJL400 and screening the plasmids in Escherichia coli by Southern hybridization using a sib‐selection technique. Sequencing the hybridizing region located an open reading frame encoding 377 amino acids. Its deduced amino acid sequence resembled that of recA genes from other bacteria. The cloned S. venezuelae gene conferred partial resistance to ethyl methanesulfonate when expressed in E. coli from the lacZ promoter.