Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Distinctive Characteristics of an Autonomous Replication Sequence of Cephalosporium acremoniurn in Yeast

Cephalosporium acremonium의 자율복제 기점의 특성

  • Lee, Kyoung (Genetic Engineering Research Institute Korea Institute of Science and Technology) ;
  • Kang, Dae-Wook (Genetic Engineering Research Institute Korea Institute of Science and Technology) ;
  • Yoon, Byung-Dae (Genetic Engineering Research Institute Korea Institute of Science and Technology) ;
  • Hwang, In-Kyu (Genetic Engineering Research Institute Korea Institute of Science and Technology) ;
  • Ahn, Jong-Seog (Genetic Engineering Research Institute Korea Institute of Science and Technology) ;
  • Min, Tae-Ick (Genetic Engineering Research Institute Korea Institute of Science and Technology)
  • 이경 (한국과학기술연구원 유전공학연구소 미생물공학연구실) ;
  • 강대욱 (한국과학기술연구원 유전공학연구소 미생물공학연구실) ;
  • 윤병대 (한국과학기술연구원 유전공학연구소 미생물공학연구실) ;
  • 황인규 (한국과학기술연구원 유전공학연구소 미생물공학연구실) ;
  • 안종석 (한국과학기술연구원 유전공학연구소 미생물공학연구실) ;
  • 민태익 (한국과학기술연구원 유전공학연구소 미생물공학연구실)
  • Published : 1991.06.01
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Abstract

An autonomous replication sequence (ARS) derived from Cephalosporium acremonium ATCC 20339 was cloned in Sarchuromyces cerevisiae SHY 3 using YIp5 as a cloning vector. A new recombinant plasmid, designated pCY-2, which contained a 3.7 kb BamHI fragment of C. acrenzonium DNA showed the highest stability among the 40 recombinant plasmids composed of the YIp5 2nd ARS of C. ucremoniztm. Also, Southern hybridization and transformation of E, cull with DNA purified from yeast transformants verified that pCY-2 autonomously replicates in yeasts. Transformation efficiency and plasmid stability of pCY-2 in yeast were higher than those ol YRp 7 containing ARS which originated from yeast. Detailed studies by subcloning revealed that two ARSs existed within 2.6 kb of the insert, which is a novel discovery. However, it was concluded that these two ARSs were ligated during the gene manipulation in vitro.

YIp5를 사용하여 Cephalosporium acremonium ATCC 2033으로부터 Saccharomyces cerevisiae SHY3에서 발현되는 자율복제 기점 (ARS)를 분리하였다. 자율복제 기점을 갖는 40종의 vector 가운데 가장 안정성 (stability)이 높은 plasmid를 pCY-2라 명명하였으며 pCY-2는 C.acremonium에서 유래하는 3.7kb의 단편을 갖고 있었다. 또 Southern hybridization과 재형질전환을 통해 pCY-2가 효모내에서 자율적으로 복제되고 있다는 것을 확인하였다. 또한 pCY-2는 형질전환율과 안정성에 있어 효모의 ARS를 갖는 YRp7 vector보다 우수하였다.

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