Improvement and Application of a Promoter-Probe Vector Bearing the PHO5 Gene as the Indicator Marker in Saccharomyces cerevisiae
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概要
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A promoter-probe plasmid vector, pVC701,in Saccharomyces cerevisiae having the repressible acid phosphatese(rAPase; EC3.1.3.2)gee, PHO5,of S. cerevisiae as the selection marker(Hwang et al., Appl. Microbiol. Biotechnol., 28,155-159,1988)was improved by inserting multiple cloning sites in the 5' non-transcribed region of PHO5 and by replacing 93-bp sequice containing three ATG sequences out of the frame but directly flanked by the translation initiation codon ATG with a 5'-CACC-3' sequence. The resulting YCp type plasmid, pVC737,is replicable in both S. cerevisiae and Escherichia coli hosts. The function of the vector was confirmed by inserting a DNA fragment bearing a promoter region of the GAP gane encoding glyceraldehyde-3-phosphate dehydrogenase of S. cerevisiae at the cloning site. Using the vector, we could analyse the cis-acting upstream regulatory region of the HIS5 gene of S. cerevisiae and also isolate Gcd-(general control derepressed) mutants occurring in the trans-acting loci of the general amino acid control.
- 社団法人日本生物工学会の論文
- 1989-01-25
著者
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Harashima Satoshi
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Harashima Satoshi
Department Of Biotechnology Faculty Of Engineering Osaka University
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Harashima Satoshi
Department Of Fermentation Technology Faculty Of Engineering Osaka University
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Hwang Y‐i
Kyungnam Univ. Masan Kor
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HWANG YONG-IL
Department of Fermentation Technology, Faculty of Engineering, Osaka University
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OSIMA YASUJI
Department of Fermentation Technology, Faculty of Engineering, Osaka University
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Osima Yasuji
Department Of Fermentation Technology Faculty Of Engineering Osaka University
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