TAT介导的白血病抑制因子受体α亚基胞内区远膜端融合蛋白的制备及其对HL-60细胞生长的抑制作用  

Preparation of TAT-mediated LIF receptor α-chain distal cytoplasmic fusion protein and its inhibitory effect against the growth of HL-60 cells

作　　者：徐莎[1] 王越[1] 孙擎[2] 刘厚奇[1] 

机构地区：[1]第二军医大学组织学与胚胎学教研室,上海200433 [2]中国人民解放军第401医院,特勤高压氧科,青岛266071

出　　处：《解剖学杂志》2014年第2期150-153,共4页Chinese Journal of Anatomy

摘　　要：目的：克隆表达HIV-1反式激活蛋白TAT与白血病抑制因子（LIF）受体α亚基胞内区C末端序列（LIFRα-CT3）的融合蛋白（TAT-CT3）,并检测其对HL-60细胞增殖的抑制作用.方法：重组构建TAT-CT3和SUMO融合表达质粒,转化大肠杆菌BL21（DE3）,IPTG诱导SUMO-TAT-CT3融合蛋白表达.表达产物经亲和层析柱纯化、去除SUMO蛋白标签后获得TAT-CT3融合蛋白,免疫印迹鉴定.采用CCK-8法检测HL-60细胞的抑制率.结果：构建了高表达重组子pET28a-SUMO-TAT-CT3,获得了相对分子质量约为26 kD的TAT-CT3融合蛋白.经活性检测,TAT-CT3融合蛋白能有效抑制HL-60细胞的生长.结论：成功地克隆、表达、纯化TAT-CT3融合蛋白,该融合蛋白具有生物活性,可抑制HL-60细胞的增殖.Objective： To prepare the TAT-CT3 fusion protein and investigate its inhibitory effect against the proliferation of HL-60 cells. Methods： The combinant plasmid harboring TAT-CT3 and SUMO gene was constructed, transformed into E. coli BL21 and induced with IPTG. The expressed fusion protein was purified by affinity chromatography and the SUMO tag was removed. The purified TAT-CT3 was detected by Western blotting. And the biological activity was tested by CCK-8. Results： pET28a-SUMO-TAT-CT3 was successfully constructed, and TAT-CT3 fusion protein with relative molecular mass about 26 kD was expressed in prokaryotic cells. Biological activity assay showed that TAT-CT3 fusion protein inhibited the growth of HL-60 cells. Conclusion： TAT-CT3 fusion protein has been successfully expressed and purified, and it can significantly inhibit the proliferation of HL-60 cells.

关 键 词：白血病抑制因子受体 TAT融合蛋白 HL-60细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]