Title Cloning and expression of the extracellular β-lactamase gene from Streptomyces sp. SMF13 in Streptomyces lividans
Author Choi, Sang Rak · Lee, Kye Joon
Address Department of Microbiology and Research Center for Molecular Microbiology, College og Natural Sciences, Seoul National University
Bibliography Korean Journal of Microbiology, 30(3),149-153, 1992
Key Words Streptomyces lividans, β-lactamase, cloning, protoplasts, β-lactam antibiotics, β-lactamase gene
Abstract Cloning of the gene encoding extracellular β-lactamase from Streptomyces sp. SMF13 in a plasmid pIJ702 and expression of the gene in Streptomyces invidans were carried out. Optimal conditions for the formation of protoplasts of S.lividans and the regeneration of the protoplasts were evaluated. Streptomyces sp. SMF-13 was selected as a donor strain of β-lactamase gene and totla DNA of the strain was partially digested with Sau3A I. DNA fragments ranged from 4kb to 10 kb were ligated to pIJ702 AT Bgl II site and then the ligated DNAs were transformed to the protoplasts of S, livivans. The transformation efficiency was 2 × 10^3 /㎍ DNA for the ligated DNA mixture. One colony among a thousand colonies regenerated showed extracellular β-lactamase and the size of the inserted DNA fragment was estimated to be 3.94 kb. The β-lactamase activity in the culture broth of the recombinant strain was maximum at 3 days culture to be 1.0 unit/ml.
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