• Title/Summary/Keyword: YEp 13 vector-B. amyloliquefaciens amylase gene cloning-hybrid DNA segregation-Expression in S.cerevosiae

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Cloning of Bacillus amyloliquefaciens amylase gene using YEp 13 as a vector II. Expression of cloned amylase gene in Saccharomyces cerevisiae (YEp 13 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning II. Saccharomyces cerevisiae에서의 발현)

  • 김관필;서정훈
    • Microbiology and Biotechnology Letters
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    • v.14 no.3
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    • pp.209-212
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    • 1986

  • $\alpha$-Amylase gene of Bacillus amyloliquetaciens was cloned on plasmid YEp13, S. cerevisiae-E. coli shuttle vector. Hybrid plasmid pTG17, carrying $\alpha$-amylase gene of B. amyloliquefaciens, was transformed to E. coli and the expression of it in yeast was investigated. This plasmid was unstable in E. coli and produced two minor plasmids, pTG17-1 and PTG17-2, which resulted from the segregation of it. Transformant of S. cerevisiae MC16 with pTG17-1 plasmid was not appeared on SD medium because of the Leu2 gene defection. S. cerevisiae could be transformed by the hybrid plasmid, and $\alpha$-amylase activity of the yeast transformant was detected by somogyi-Nelson method and agar diffusion method.

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