| 人血管内皮抑制素基因在毕赤甲醇酵母中的表达 |
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| 引用本文： | 曹志飞,李新平,潘春娟,邱玲娅.人血管内皮抑制素基因在毕赤甲醇酵母中的表达[J].苏州科技学院学报(自然科学版),2005,22(4):57-60,72. |
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| 作者姓名： | 曹志飞 李新平 潘春娟 邱玲娅 |
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| 作者单位： | 苏州大学,生命科学学院,江苏,苏州,215006 |
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| 摘 要： | 对发生突变的人血管内皮抑制素基因进行了PCR修正,并将其连接到pAO815载体上,然后克隆进大肠杆菌TOP10F＇中,提取质粒测序,证实序列正确.再用电激法转化毕赤甲醇酵母KM71,利用RDB平板和PCR技术筛选出阳性克隆菌落后,甲醇诱导表达,SDS-PAGE电泳检测显示重组的人血管内皮抑制素蛋自在毕赤甲醇酵母KM71中获得了有效表达.
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| 关 键 词： | 人血管内皮抑制素 PCR定点突变 毕赤甲醇酵母 pAO815载体 诱导表达 |
| 文章编号： | 1672-0687（2005）04-0057-04 |
| 收稿时间： | 2005-04-26 |
| 修稿时间： | 2005-04-26 |
Successful Expression of Human Endostatin Gene in Pichia pastoris |
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| CAO Zhi-fei,LI Xin-ping,PAN Chun-juan,QIU Ling-ya.Successful Expression of Human Endostatin Gene in Pichia pastoris[J].Journal of University of Science and Technology of Suzhou,2005,22(4):57-60,72. |
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| Authors: | CAO Zhi-fei LI Xin-ping PAN Chun-juan QIU Ling-ya |
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| Institution: | Dept. of Cellular and Molecular, Suzhou University, Suzhou 215006, China |
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| Abstract: | The mutant human endostatin modified by PCR,is ligated into pAO815 vector and cloned into E.coli TOP10F'.The sequence of this modified gene is proved to be correct by measuring the sequence of the collected particles.Then the isolated plasmid pAO815-endostatin is transformed into Pichia pastoris KM71 by electroporation.After the positive recombinants are screened out by using RDB plate and PCR technique and induced with methanol,the SDS-PAGE results show that human endostatin protein is effectively expressed in Pichia pastoris KM71. |
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| Keywords: | human endostatin PCR site-directed mutagenesis Pichia pastoris pAO815 vector induced expression |
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