融合蛋白TAT-p53-MTS的促细胞凋亡作用  

Pro-apoptotic effect of the recombinant protein TAT-p53-MTS

作　　者：吴兰[1] 赵梓亦[1] 李灵[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2015年第1期170-174,共5页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金(31000579)

摘　　要：本研究首先构建了融合蛋白TAT-p53-MTS的原核表达质粒pET-22b(+)-TATp53-MTS,将表达纯化的融合蛋白与p53缺陷型人肺癌细胞NCI-H1299孵育,通过Western blot和免疫共沉淀(co-immunoprecipitation,co-IP)检测融合蛋白的细胞穿膜以及与Bcl-2蛋白的相互作用,并通过流式细胞术检测融合蛋白对NCI-H1299细胞的促凋亡作用.结果显示,融合TAT和MTS的p53重组蛋白具有更强的细胞穿膜及线粒体定位能力;并且,融合蛋白能够通过与Bcl-2蛋白的结合显著诱导NCI-H1299细胞的凋亡.The plasmid for prokaryotic production of recombinant p5 3 fused with TAT and MTS （TAT-p5 3-MTS）was constructed.Purified recombinant protein was incubated with p5 3-deficient human lung cancer line NCl-H1299,and the cell lysate was subjected to co-immunoprecipitation and Western blot for testing the trans-membrane ability of the fusion protein,as well as the interaction between the fusion protein and Bcl-2.Additionally,flow cytometry was employed to detect fusion protein’s proapoptotic impact on NCl-H1299 cells.The results showed that TAT and MTS can improve both the trans-mem-brane ability and the mitochondrial targeting of the fusion protein.What’s more,the fusion protein ex-hibits ability to interact with Bcl-2,through which apoptosis is significantly induced.

关 键 词：P53 细胞凋亡 线粒体定位 MTS TAT 

分 类 号：Q955[生物学—动物学]