| 人源血管紧张素转化酶-N结构域基因片段在毕赤酵母中的表达 |
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| 引用本文： | 徐辉,徐珏,陈成集,丁明,许传莲. 人源血管紧张素转化酶-N结构域基因片段在毕赤酵母中的表达[J]. 浙江理工大学学报, 2011, 28(4) |
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| 作者姓名： | 徐辉 徐珏 陈成集 丁明 许传莲 |
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| 作者单位： | 浙江理工大学生命科学学院,杭州,310018 |
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| 摘 要： | 利用RT-PCR方法从食管癌细胞TE-1中反转出cDNA,PCR扩增得到血管紧张素转化酶N结构域(ACE-N)基因片段,构建pPIC9K-ACE-N表达载体,转化毕赤酵母GSll5,阳性克隆再次电转,并从转化菌株中筛选出多拷贝转化子进行表达.经Ni2+亲和层析柱对表达的蛋白进行纯化,获得的目的蛋白表达量为0.11 g/L,其纯度大于99％.为今后选择性抑制血管紧张素转化酶两个同源结构域的体外研究奠定基础.
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| 关 键 词： | 血管紧张素转化酶-N结构域 RT-PCR 毕赤酵母 |
Expression of Human Angiotensin Converting Enzyme-N Domain in Pichia Pastoris |
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| XU Hui,XU Jue,CHEN Cheng-ji,DING Ming,XU Chuan-lian. Expression of Human Angiotensin Converting Enzyme-N Domain in Pichia Pastoris[J]. Journal of Zhejiang Sci-tech University, 2011, 28(4) |
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| Authors: | XU Hui XU Jue CHEN Cheng-ji DING Ming XU Chuan-lian |
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| Affiliation: | XU Hui,XU Jue,CHEN Cheng-ji,DING Ming,XU Chuan-lian(School of Life Sciences,Zhejiang Sci-Tech Universtry,Hangzhou 310018,China) |
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| Abstract: | The authors obtain the cDNA by RT-PCR from the TE-1 cell line,derived ACE-N gene fragment through PCR,and construct secretory expression plasmid pPIC9K-ACE-N.The recombinant plasmid is transformed into Pichia pastoris strain GS115,and the positive clones were selected and subjected to electroporation.Antibiotic G418 is used for screening multi-copy inserts.The expressed protein is purified by Ni2+ affinity chromatography.The expression quantity of target protein reached 0.11g/L and its purity reaches 99%.Th... |
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| Keywords: | angiotensin converting enzyme-N domain RT-PCR pichia pastoris |
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