• Title/Summary/Keyword: B.amyloliquefaciens-Amylase gene clonig-YEp13-Expression in E.coli

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Cloning of Bacillus amyloliquefaciens amylase gene using YEp13 as a vector I. Expression of cloned amylase gene in Escherichia coli (YEp 13 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning I. Escherichia coli에서의 발현)

  • 이창후;서정훈
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.155-160
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    • 1986

  • $\alpha$-Amylase gene of B. amyloliquefaciens was cloned to E. coli-yeast shuttle vector YEp-13 and expressed in E. coli. Chromosomal DNA of B. amyloliquefaciens was partially digested with Sau3Al and YEp13 plasmid was cleaved with BamH1. The hybrid plasmid, pHA28, was constructed by shotgun method and transformed to E. coli C600 and HB101. The amount of $\alpha$-amylase produced by transformants of E. coli was about 20% to 30% of that produced by B. amyloli-quefaciens. About 65% of $\alpha$-amylase produced by transformant was secreted into periplasm and the others were located in cytoplasm. $\alpha$-Amylase production was maximal when transformants were cultivated for 15hr to 20hr. As the result of agarose gel electrophoresis, pHA28 plasmid was found to be various in its size. This result suggested that pHA28 plasmid was segregated.

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