High Level Expression of Human Transforming Growth Factor α Gene in Escherichia coli in a Fusion Gene System with Human Growth Hormone Gene
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概要
- 論文の詳細を見る
An artificial gene encoding[Leu^<14>]human growth hormone(hGH), which had a leucine residue at position 14 instead of methionine as in the wild type hGH protein, was constructed and expressed in Escherichia coli at a high level, as well as the[Met^<14>]hGH gene, under the control ot the trp promoter including a consensus SD sequence. A human transforming growth factor α(TGFα)gene, which had a methionine codon attached to the 5'-terminal region, was chemically synthesized and fused to the[Lue^<14>]hGH gene. THe fusion gene was efficiently expressed in E. coli, yielding 48mg per gram of the wet cell weight. After the fusion protein was cleaved into two parts at the single methionine site by treatment with cyanogen bromide, the desired TGFα was made to form disulfide bonds by a refolding reaction, and then purified by HPLC. The structure of the TGFα was confiremed by amino acid sequence analysis. The biological activity of the purified TGFα was assessed.
- 社団法人日本生物工学会の論文
- 1991-04-25
著者
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Miyake Tetsuo
Institute For Biotechnology Research Wakunaga Pharmaceutical Co. Ltd.
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Sugiyama M
Institute Of Pharmaceutical Sciences Hiroshima University School Of Medicine
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Sugiyama M
Hiroshima Univ. Hiroshima Jpn
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Sugiyama Masanori
Institute Of Pharmaceutical Sciences Hiroshima University School Of Medicine
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MISOKA FUSAKAZU
Institute for Biotechnology Research, Wakunaga Pharmaceutical Co. Ltd.,
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SAKAMOTO SHUNJI
Institute for Biotechnology Research, Wakunaga Pharmaceutical Co., Ltd.,
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Sugiyama Masanori
Institute Of Pharmaceutical Sciences Faculty Of Medicine Hiroshima University
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Sakamoto S
Institute For Biotechnology Research Wakunaga Pharmaceutical Co. Ltd.
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Sakamoto Shunji
Institute For Biotechnology Research Laboratories Wakunaga Pharmaceutical Co. Ltd.
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Misoka Fusakazu
Institute For Biotechnology Research Wakunaga Pharmaceutical Co. Ltd. :(present Address)technology C
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