Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Studies on the Development of Yeast Promoter for the Gene Expression

효모(酵母) 유전자(遺傳子) 발현용(發現用) Promoter 개발(開發)에 관(關)한 연구(硏究)

  • Chung, Ho-Kwon (Department of Microbial Technology, Kon-Kuk University) ;
  • Park, Joon-Hee (Department of Food Science and Technology, Chung-Ang University) ;
  • Shim, Sang-Kook (Department of Food Technology, Dongnam Health Junior College) ;
  • Chung, Dong-Hyo (Department of Food Science and Technology, Chung-Ang University)
  • 정호권 (건국대학교 공과대학 미생물학과) ;
  • 박준희 (중앙대학교 산업대학 식품가공학과) ;
  • 심상국 (동남보건전문대학 식품가공과) ;
  • 정동효 (중앙대학교 산업대학 식품가공학과)
  • Published : 1995.02.28
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Abstract

The purpose of this study was the development of promoter for the lacZ' gene. Two heterologous promoter I and II of lacZ' gene were isolated from chromosomal DNA Bam HI fragment of yeast. The size of the promoter I was estimated to be 2.5 kb and ${\beta}-galactosidase$ activity was 124.6 U/mg protein, and the size of the promoter II was 4.0 kb and its ${\beta}-galactosidase$ activity was 168.8 U/mg protein, respectively. The stability of the recombinant YEp plasmid in the transformant was from 52.7 to 67.4% at minimal medium. YIp plasmid was constructed from YEp plasmid, and expressed both in E. coli and yeast. The promoter I aid II iso-lated from yeast chromosomal DNA can be used for promoter of plasmid YEp and YIp.

본 연구는 lacZ' 유전자의 promoter 개발을 위하여 착수하였다. lacZ' 유전자의 heterologous promoter I과 II를 효모 염색체의 Bam HI DNA 단편에서 분리하였다. Promoter I의 크기는 2.5 Kb 정도이고 ${\beta}-galactosidase$ 활성은 124.6 U/mg protein이었으며 promoter II의 크기와 효소활성은 4.0 Kb와 168.8 U/mg이었다. 형질 전환체에서의 YEp plasmid 안정성은 52.7%에서 67.4% 정도였다. YEp plasmid로부터 YIp plasmid를 재조합하였으며 이 YIp plasmid는 대장균에서나 효모에서도 발현되었다. 효모로부터 분리한 promoter I과 II는 재조합된 YEp와 YIp plasmid의 promoter로서 이용 가능하였다.

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