| 凋亡素融合蛋白的基因克隆、表达与体外活性分析 |
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| 引用本文： | 赵健,高鹏,李素霞,袁勤生,刘建文.凋亡素融合蛋白的基因克隆、表达与体外活性分析[J].华东理工大学学报(自然科学版),2009,35(2). |
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| 作者姓名： | 赵健 高鹏 李素霞 袁勤生 刘建文 |
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| 作者单位： | 华东理工大学生物反应器工程国家重点实验室,上海,200237 |
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| 摘 要： | 来源于HIV-1病毒(47~57位氨基酸)的TAT小肽具有跨膜功能,可以将外源蛋白进行跨膜转运。通过聚合酶链反应(PCR)的方法扩增了TAT-凋亡蛋白序列,与载体pET-28b连接后在大肠杆菌BL21(DE3)中获得了高表达,以包涵体形式表达的TAT-凋亡蛋白在变性条件下进行了Ni-NTA纯化,纯化的蛋白经MTT法证明具有诱导HeLa细胞凋亡的能力。
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| 关 键 词： | TAT-apoptin over-expression purification MTT（3-（4 5-Dimethylthigal-2-yl）-2 5-（diphenyltetragalium） bromide） |
Cloning and Expression of TAT-apoptin and Its Activity Study in vitro |
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| ZHAO Jian,GAO Peng,LI Su-xia,YUAN Qin-sheng,LIU Jian-wen.Cloning and Expression of TAT-apoptin and Its Activity Study in vitro[J].Journal of East China University of Science and Technology,2009,35(2). |
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| Authors: | ZHAO Jian GAO Peng LI Su-xia YUAN Qin-sheng LIU Jian-wen |
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| Institution: | State Key Laboratory of Bioreactor Engineering;East China University of Science and Technology;Shanghai 200237;China |
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| Abstract: | A basic peptide derived from human immunodeficiency virus HIV-1 TAT peptide(positions 47-57) can translocate through the cell membranes,the characteristics of which are utilized for the deli-(very) of exogenous proteins into cells. The coding sequence for the TAT-apoptin was amplified by polymerase chain reaction(PCR),then inserted into pET28b vector and highly expressed in Escherichia coli BL21(DE3).The protein expressed in inclusion body was purified by Ni-NTA under denaturing condition.(Apoptosis) activi... |
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