Molecular cloning of phospho-beta-galactosidase gene of Lactobacillus casei in Escherichia Coli
1989
Moon, K.H. | Park, C.H. | Choi, S.Y. | Lee, Y.M. | Min, K.H. (Sookmyung Women's Univ., Seoul (Korea R.). Dept. of Biology) | Kim, T.H. (Ildong Pharm Company Limited, Seoul (Korea R.). Central Laboratory) | Kim, Y.S. (Seoul National Univ., Seoul (Korea R.). Coll. of Natural Sciences)
Gene for lactose catabolism in Lactobacillus casei SW-M1 was encoded by a 60Kb metabolic plasmid. A derivative of only 10kb, pPLac 15 of recombinant plasmid, was constructed by introducing into pBR322 and was cloned into E. coli using restriction endonuclease Pst I. A 10kb insert DNA in plasmid pBR322 was identified as a gene encoded phospho-beta-galactosidase by the determination of enzyme activity. Phospho-beta-galactosidase was apparently expressed in E. coli. The enzyme activities of cell-free extract from transformant E. coli HB101 carrying pPLac 15 DNA were not different from that of L. casei as a donor strain on the basis of enzyme properties. However specific activity of phospho-beta-galactosidase in the cloned strain with Lac Y** (-) phenotype of E. coli HB101 was lower than that in L. casei strain
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