Construction of Inducible Secretion Vectors and Their Application for the Secretion of Foreign Extracellular and Intracellular Proteins in Bacillus subtilis
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概要
- 論文の詳細を見る
We have constructed inducible secretion vector (pISA412) which has (i) a unique BamHI site as a cloning site between the signal sequence and transcriptional terminator of Bacillus subtilis α-amylase gene (amyE), and (ii) penI-penJ operon and promoter-operator regions of penP (penicillinase gene) from Bacillus licheniformis as a regulation system upstream to the amyE signal sequence. Using pISA412,we examined the expression in B. subtilis of several cloned secretory protein genes such as the penicillinase gene from B. licheniformis (penP), α-amylaes genes from Bacillus stearothermophilus (amyT) and human salivary (hsa), and the thermostable endoglucanase gene from Clostridium thermocellum (engD). In each case, gene expression was induced by the addition of 2-(2'-carboxyphenyl) benzoyl-6-aminopenicillanic acid (CBAP). Significant levels of extracellular activity of these gene products could be detected. Furthermore, when an intracellular enzyme gene (β-galactosidase gene of Escherichia coli; lacZ) was subcloned in the secretion vector, β-galactosidase activity could be detected in the culture supernatnat. We also constructed a secretion vector, pISA^Χ<ts>412,with the temperaturesensitive repressor (P70L) in order to induce gene expession simply by shifting temperature of the culture from 30℃ to 48℃. Expression of the cloned gene into the pISA^<ts>412 could be controlled by the temperature shift. These results indicate that pISA412 and pISA^<ts>412 vectors are useful inducible secretion vectors in B. subtilis.
- 公益社団法人日本生物工学会の論文
- 1993-07-25
著者
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Shima Hideki
The Graduate School Of Engineering Osaka University:(present Address)texas Instruments Japan Ltd.
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TAKAGI MASAHIRO
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Shima Hideki
Department Of Biotechnology Faculty Of Engineering Osaka Univesity
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Imanaka Tadayuki
Department Of Applied Biotechnology Faculty Of Engineering Osaka University
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Bhatnagar Lakshmi
Michigan Biotechnology Institute
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Zeikus J.gregory
Michigan Biotechnology Institute
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Shima Hideki
Department Of Applied Physics Graduate School Of Engineering Yokohama National University
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Imanaka Tadayuki
Department Of Biotechnology Faculty Of Engineering Osaka Univesity
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Takagi Masahiro
Department Of Biotechnology Faculty Of Engineering Osaka University
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Takagi Masahiro
Department Of Biotechnology Faculty Of Engineering Osaka Univesity
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