| Secretory expression of α single-chain insulin precursor in yeast and its conversion into human insulin |
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| 引用本文： | 张友尚,胡红明,蔡若蓉,冯佑民,朱尚权,贺潜斌,唐月华,徐明华,许英镐,张新堂,刘滨,梁镇和.Secretory expression of α single-chain insulin precursor in yeast and its conversion into human insulin[J].中国科学：生命科学英文版,1996(3). |
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| 作者姓名： | 张友尚 胡红明 蔡若蓉 冯佑民 朱尚权 贺潜斌 唐月华 徐明华 许英镐 张新堂 刘滨 梁镇和 |
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| 作者单位： | State Key Laboratory of Molecular Biology，Shanghai Institute of Biochemistry，Chinese Academy of Sciences，Shanghai 200031，China |
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| 基金项目： | Project supported by the National High Technology Program of China. |
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| 摘 要： | A synthetic single-chain porcine insulin precursor (PIP) gene and an α-mating factor leader sequence (αMFL) gene obtained by the PCR method are inserted between the promoter and 3'-terminating sequence of the alcohol dehydrogenase gene ADH1 in plasmid pVT102-U to form plasmid pVT102-U/α MFL-PIP. The single-chain insulin precursor is expressed and secreted to the culture medium by Saccharomyces cererisiae transformed by pVT102-U/αMFL-PIP. The precursor is purified and converted into human insulin by tryptic transpeptidation. The purified human insulin is fully active and can be crystallized. The overall yield of human insulin is 25 mg per liter of culture medium.
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