Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Cloning and Expression of Thermostable Alpha-amylase Gene in Escherichia coli from Bacillus licheniformis ATCC 27811

Bacillus licheniformis ATCC 27811이 생산하는 내열성 $\alpha$-amylase 유전자의 Cloning 및 발현

  • Kim, I.C. (Department of Food Science and Technology, Seoul National University) ;
  • Jang, S.Y. (Department of Food Science and Technology, Seoul National University) ;
  • Cha, J.H. (Department of Food Science and Technology, Seoul National University) ;
  • Ko, Y.H. (Department of Food Science and Technology, Seoul National University) ;
  • Park, K.H. (Department of Food Science and Technology, Seoul National University) ;
  • Rho, H.M. (Department of Zoology, Seoul National University)
  • 김인철 (서울대학교 식품공학과) ;
  • 장소영 (서울대학교 식품공학과) ;
  • 차재호 (서울대학교 식품공학과) ;
  • 고영환 (서울대학교 식품공학과) ;
  • 박관화 (서울대학교 식품공학과) ;
  • 노현모 (서울대학교 동물학과)
  • Published : 1988.10.01
Download PDF
⟨ Previous Next ⟩

Abstract

The gene for thermostable alpha-amylase from the thermostable bacterium Bacillus licheniformis has been cloned and expressed in Escherichia coli. The Alpha-amylase producing E. coli cells contained a 7.4 kb chimeric plasmid (pTA 322) which was composed of the vector pBR322 and a 3.1 kb EcoRI fragment of B. licheniformis DNA. The alpha-amylase from cloned fragement was shown to be indistlnguishable from that of B. licheniformis in the optimum temperature of 9$0^{\circ}C$, heat stability and the pH stability. The foreign gene was expressed efficiently in E. coli and stably maintained.

Bacillus licheniformis ATCC 27811의 염색체 DNA를 분리하며 제한효소인 EcoRI으로 부분절단하고, 동일 효소로 절단한 plasmid pBR322에 ligation 시킨 뒤 E. coli HB 101에 형질전환시켜 alpha-amylase 형질을 보여주는 균주를 선별하였다. 선별된 형질전환체로부터 alpha-amylase를 분리하며, 원 균주인 Bac. licheniformis가 생산하는 alpha-amylase와 pH 및 온도특성을 비교하며 모균주와 같은 성질을 가졌음을 확인하였다. 재조합체 DNA로부터 얻은 Insert는 대략 3.1kb 정도였고, HindIII, ClaI, PstI, SalI Site를 한개씩 가지고 있었다.

Keywords