人端粒逆转录酶与Snapin蛋白相互作用的鉴定

摘要

目的:研究人端粒逆转录酶(hTERT)与Snapin蛋白的相互作用.方法:将hTERT基因和Snapin基因分别构建到pGBKT7和pGADT7载体中,用酵母双杂交方法在酵母中验证其相互作用；在293T细胞中,共转染带有Flag标签的hTERT及带有GFP标签的Snapin质粒,进行免疫共沉淀；将GST融合的Snapin纯化蛋白与hTERT进行GST-pull down,验证其相互作用.结果:3种方法都证明hTERT能够与Snapin相互作用.结论:Snapin蛋白能够与hTERT相互作用,为研究Snapin参与调控端粒酶的功能活动提供了一些线索.%Objective: To identify the interaction of human telomere reverse transcriptase (hTERT) and Snapin. Methods: The hTERT gene and Snapin gene were constructed into pGBKT7 and pGADT7 vector respectively, the yeast two-hybrid technique was used to confirm their interaction in yeast cells. The flag tagged hTERT and GFP fused Sanpin vectors were contransfected into 293T cells, and immunoprecipitation and GST-pull down were performed to confirm the interaction. Results: Confirmed by the three techniques, Snapin can interact with hTERT. Conclusion: The interaction between the hTERT and Snapin which provides clues for the research of Snapin as a regulator in the telomerase functioning pathway.