Molecular Cloning of Serratia rnarcescens Metalloprotease Gene into Escherichia coli

Serratia marcescens Metalloprotease 유전자의 대장균에로의 클로닝

  • 김기석 (경상대학교 자연과학대학 미생물학과) ;
  • 이창원 (경상대학교 자연과학대학 미생물학과) ;
  • 이상열 (경상대학교 미생물학과) ;
  • 이병룡 (코오롱주식회사) ;
  • 신용철 (경상대학교 자연과학대학 미생물학과)
  • Published : 1992.06.01

Abstract

Molecular cloning of metalloprotease gene from Serratia marcescens ATCC 21074 into Escherichia coli JM109 was carried out. Chromosomal DNA of S. marcescens was completely digested with Hind111 and southern hybridization with a synthetic oligonucleotide probe revealed that a 50 KD metalloprotease gene was contained in 4.0 Kb chromosomal DNA fragment, 4.0 Kb chromosomal DNA fragments eluted from agarose gel were ligated with pUC19 and transformed into E. coli JM109. Nine positive clones were obtained from about $1\times 10^3$ transformants by colony hybridization. Their recombinant plasmids, pSPl and pSP2 have same chromosomal DNA fragments in pUC19 in opposite-orientations. When cloned metalloprotease gene was expressed in E. coli, about 52 KD precursor protein of metalloprotease was detected by western blot analysis from E. coli harboring a recombinant plasmid pSP2. Plasmid pSP2 showed no protease activities in E. coli but overproduced the active metalloprotease in S. rnarcescens ATCC 27117.

Keywords

Serratia marcescens;cloning of 50KD metalloprotease gene;expression of metalloprotease