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    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, (2): 202-206.
    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, (2): 202-206.
    Cloning and Expression of TATapoptin and Its Activity Study in vitro[J]. Journal of East China University of Science and Technology, 2009, (2): 202-206.
    Citation: Cloning and Expression of TATapoptin and Its Activity Study in vitro[J]. Journal of East China University of Science and Technology, 2009, (2): 202-206.

    凋亡素融合蛋白的基因克隆、表达与体外活性分析

    Cloning and Expression of TATapoptin and Its Activity Study in vitro

    • 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。

       

      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 activity of the TAT apoptin was evaluated by MTT. Results indicated that HeLa cells were highly sensitive to the purified TAT apoptin.

       

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