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Expression of Mouse $alpha-Amylase$ Gene in Methylotrophic Yeast Pichia pastoris
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  • Expression of Mouse $alpha-Amylase$ Gene in Methylotrophic Yeast Pichia pastoris
  • Expression of Mouse $alpha-Amylase$ Gene in Methylotrophic Yeast Pichia pastoris
저자명
Uehara. Hiroyuki,Choi. Du Bok,Park. Enoch Y.,Okabe. Mitsuyasu
간행물명
Biotechnology and bioprocess engineering
권/호정보
2000년|5권 1호|pp.7-12 (6 pages)
발행정보
한국생물공학회
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정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

The expression of the mouse $alpha-amylase$ gene in the methylotrophic yeast, P pastoris was investigated. The mouse $alpha-amylase$ gene was inserted into the multi-cloning site of a Pichi a expression vector, pPIC9, yielding a new expression vector pME624. The plasmid pME624 was digested with SalI or BglII, and was introduced into P. pastoris strain GSl15 by the PEG1000 method. Fifty-three transformants were obtained by the transplacement of pME624 digested with SaiII or BglII into the HIS4locus $(38;of;Mut^+;clone)$ or into the AOX1 locus $(15;of;Mut^s;clone)$. Southern blot was carried out in 11 transformants, which showed that the mouse $alpha-amylase$ gene was integrated into the Pichia chromosome. When the second screening was performed in shaker culture, transformant G2 showed the highest $alpha-amylase$ activity, 290 units/ml after 3-day culture, among 53 transformants. When this expression level of the mouse $alpha-amylase$ gene is compared with that in recombinant Saccharomyces cerevisiae harboring a plasmid encoding the same mouse $alpha-amylase$ gene, the specific enzyme activity is eight fold higher than that of the recombinant S. cerevisiae.