PEP-1-VP3融合蛋白的分离纯化与初步鉴定
Prokaryotic expression and purification of PEP-1-VP3 fusion protein
作者：蔡召忠;陈萍;邓守恒;
Author：
收稿日期： 年卷（期）页码：2015,30(02):-160-162
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：PEP-1-VP3融合蛋白;分离;纯化;诱导表达;克隆;鉴定
Key words：
基金项目：湖北省卫生厅基金(QJX2008-42);;
湖北医药学院创新基金(2008CXZ02,2010XSA02);;
十堰市科技局基金(2010st24)
中文摘要
目的构建融合蛋白PEP-1-VP3的原核表达质粒,并进行诱导表达和蛋白纯化。方法通过PCR方法自PGEX-6P-1/TAT-VP3质粒中扩增VP3基因,运用靶向克隆酶,将VP3基因插入到线性载体p ET15b-PEP-1中,构建重组表达质粒p ET15b-PEP-1-VP3,经测序证实构建成功后转化E.coli Rosetta,表达PEP-1-VP3融合蛋白,经Ni-NTA树脂亲和层析,进行SDS-PAGE及Western Blot分析鉴定。结果成功构建PEP-1-VP3融合蛋白原核表达载体,表达并纯化了17.6k D的PEP-1-VP3融合蛋白。结论所构建的PEP-1-VP3融合蛋白为体内抗肿瘤研究提供了理论基础。
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