CAJ | 학술논문

目的：应用酵母双杂交系统筛选与端粒重复序列结合蛋白1（ TRF1）相互作用的蛋白质，探讨TRF1在HeLa细胞有丝分裂期的调控机制。方法：应用酵母双杂交技术，以pGBKT7-TRF1为诱饵质粒筛选人HeLa细胞cDNA文库，并对筛选结果进行测序及同源性分析。运用免疫共沉淀、体外沉降实验验证SAM68与TRF1是否能够相互结合。结果：酵母双杂交结果共发现5个可能与TRF1存在相互作用的蛋白，包括人SAM68蛋白、Cullin1蛋白、Plk1蛋白、Hsp70蛋白以及ATF5蛋白。体内免疫共沉淀实验和体外沉降实验证实SAM68能够与TRF1在体内与体外相互结合。结论：SAM68可能是一个新的TRF1结合蛋白，TRF1有可能参与了有丝分裂期的调控。
목적：응용효모쌍잡교계통사선여단립중복서렬결합단백1（ TRF1）상호작용적단백질，탐토TRF1재HeLa세포유사분렬기적조공궤제。방법：응용효모쌍잡교기술，이pGBKT7-TRF1위유이질립사선인HeLa세포cDNA문고，병대사선결과진행측서급동원성분석。운용면역공침정、체외침강실험험증SAM68여TRF1시부능구상호결합。결과：효모쌍잡교결과공발현5개가능여TRF1존재상호작용적단백，포괄인SAM68단백、Cullin1단백、Plk1단백、Hsp70단백이급ATF5단백。체내면역공침정실험화체외침강실험증실SAM68능구여TRF1재체내여체외상호결합。결론：SAM68가능시일개신적TRF1결합단백，TRF1유가능삼여료유사분렬기적조공。
Aim:To screen and identify the interactive proteins of TRF 1 and provide clues for the studies on TRF 1 func-tion in mitosis.Methods:The human HeLa cell cDNA library was screened with pGBKT 7-TRF1 as bait plasmid by yeast two-hybrid system, and positive yeast clones were obtained , which were sequenced and analyzed with Blast in NCBI .The in vivo immunoprecipitation and the in vitro pulldown experiments were used to identify the interaction between SAM 68 and TRF1.Results:Five proteins,human SAM68,Cullin1,Plk1,Hsp70 and ATF5,that may interact with TRF1 were found.In vivo immunoprecipitation and in vitro pulldown experiments showed that SAM 68 could interact with TRF1.Conclusion:SAM68 is a novel TRF1 interacting protein;TRF1 may play an important role in the functional regulation of mitosis .