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利用酵母双杂交系统在Hela cDNA文库中筛选与18型人乳头瘤病毒E6癌基因相互作用的新型结合蛋白

Screening for novel binding proteins interacting with human papillomavirus type 18 E6 oncogene in the Hela cDNA library by yeast two-hybrid system.

作者信息

Li Shuang, Liu Ping, Xi Ling, Jiang Xuefeng, Zhou Jianfeng, Wang Shixuan, Meng Li, Lu Yunping, Ma Ding

机构信息

Cancer Biology Center, Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.

出版信息

J Huazhong Univ Sci Technolog Med Sci. 2008 Feb;28(1):93-6. doi: 10.1007/s11596-008-0124-7.

Abstract

To screen for novel binding proteins interacting with high-risk HPV 18 E6 oncogene, the strain AH109 was transformed with pGBKT7-HPV18 E6 plasmid, and subsequent transference was utilized to screen for interacting proteins with HPV 18 E6 in human Hela cDNA library. HPV18 E6 mRNA was expressed in yeast and there was no self-activation and toxicity in AH109. Seven proteins that interacted with HPV18 E6, including transmembrane protein 87B, phosphonoformate immuno-associated protein 5, vimentin, KM-HN-1 protein, dedicator of cytokinesis 7, vaccinia related kinase 2 and a hypothetical protein, were identified. It was suggested that yeast two-hybrid system is an efficient for screening interacting proteins. The high-risk HPV 18 E6 oncogene may interact with the proteins, which may be associated with signal transduction and transcriptional control, epithelial cell invasion and migration, as well as humoral and cellular immune etc. This investigation provides functional clues for further exploration of potential oncogenesis targets for cancer biotherapy.

摘要

为筛选与高危型人乳头瘤病毒18型(HPV 18)E6癌基因相互作用的新型结合蛋白,将AH109菌株用pGBKT7-HPV18 E6质粒进行转化,随后利用其在人Hela cDNA文库中筛选与HPV 18 E6相互作用的蛋白。HPV18 E6 mRNA在酵母中表达,且AH109无自激活和毒性。鉴定出7种与HPV18 E6相互作用的蛋白,包括跨膜蛋白87B、膦甲酸免疫相关蛋白5、波形蛋白、KM-HN-1蛋白、胞质分裂 dedicator 7、痘苗相关激酶2和一种假定蛋白。提示酵母双杂交系统是筛选相互作用蛋白的有效方法。高危型HPV 18 E6癌基因可能与这些蛋白相互作用,这些蛋白可能与信号转导和转录调控、上皮细胞侵袭和迁移以及体液和细胞免疫等有关。本研究为进一步探索癌症生物治疗潜在的致癌靶点提供了功能线索。

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