HeLa细胞端粒酶的分离纯化及其蛋白质组分的研究

The Study of Purification of HeLa Cells Telomerase and Its Protein Components.

根据端粒酶含有蛋白质组分和RNA组分的特点 ,采用寡核苷酸亲和纯化法从HeLa细胞蛋白粗提物中分离纯化人类端粒酶 ,纯化产物以TRAP法检测其延伸端粒活性 ,并采用RNA印迹法进行鉴定 ,然后从纯化产物中分离蛋白质组分 ,以SDS 聚丙烯酰胺凝胶电泳检测其蛋白质亚基成分 ,可见到 4种蛋白质亚基成分 ,与蛋白质分子质量标准比较 ,有两条位置接近 2 12 2ku ,一条接近 116 0ku ,一条接近 42 7ku .结果表明 ,蛋白质寡核苷酸亲和纯化法一步性分离纯化HeLa细胞端粒酶可得到端粒酶活性片段 .

To purify the telomerase of HeLa cells and to study its protein components. The method of affinity purifying was used to extract the telomerase component from the crude extract of HeLa cells, basing on the specificity of telomerase containing protein and RNA. And then the activity of telomerase was tested, SDS PAGE was used to test protein components . There four bands were observed in the SDS PAGE gel, there are two near 212 2 ku, one near 97 4 ku, one near 42 7 ku compared with the protein mark. It was shown that the product with telomerase activity was obtained using the affinity purification method.