Open AccessMolecular cloning of the genes for anthranilate synthetase from Streptomyces venezuelae ISP 5230
Author(s) -
Paradkar Ashish S.,
Stuttard Colin,
Vining Leo C.
Publication year - 1991
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.1574-6968.1991.tb04439.x
Subject(s) - cloning (programming) , gene , biology , molecular cloning , streptomyces , genetics , computational biology , bacteria , gene expression , computer science , programming language
Summary Fragments of genomic DNA from Streptomyces venezuelae ISP5230 were cloned in the Escherichia coli expression vector pTZ18R and the plasmids were used to trasform E. coli JA194 ( trpE ). The transformants included a prototrophic strain containing a recombinant plasmid, pDQ181, with an approximately 6.8‐kb insert. Subcloning located the trpE ‐complementing DNA in a 2.4‐kb segment. Transformation of E. coli ED23 (lacking both trpE and trpG functions) with plasmids containing the 2.4‐kb DNA segment gave prototrophic strains exhibiting both the ASI and ASII activities of anthranilate synthetase. The results indicated that trpE and trpG are clustered in S. venezuelae . Regions hybridizing to the pDQ181 insert were present in the genomic DNA of other streptomycetes.