Cytochrome C oxidase III interacts with hepatitis B virus X protein in vivo by yeast two-hybrid system
Cytochrome C oxidase III interacts with hepatitis B virus X protein in vivo by yeast two-hybrid system作者机构：DepartmentofGastroenterologyRenminHospitalofWuhanUniversityWuhan430060HubeiProvinceChina DepartmentofGastroenterologyUnionHospitalofFujianMedicalUniversityFuzhou350001FujianProvinceChina ResearchInstituteofHepatologyBeijingUniversityBeijing100044China
出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)
年 卷 期：2004年第10卷第19期
页 面：2805-2808页
核心收录：
学科分类：1004[医学-公共卫生与预防医学(可授医学、理学学位)] 100401[医学-流行病与卫生统计学] 10[医学]
基 金：Supported by Science and Technology Program of Fujian Province No.2001Z157
主 题：细胞色素C 氧化酶III HBV 乙型肝炎病毒 X蛋白质 体内 酵母 HBV 二混合物系统
摘 要：AIM: To screen and identify the proteins which interact with hepatitis B virus (HBV) X protein in hepatocytes by yeast two-hybrid system and to explore the effects of X protein in the development of hepatocellular carcinoma (HCC).METHODS: With HBV X gene amplified by polymerase chainreaction (PCR), HBV X bait plasmid, named pAS2-1-X, was constructed by yeast-two hybridization system3 and verified by auto-sequencing assay, pAS2-1-X was transformed into the yeast AH109, and X-BD fusion protein expressed in the yeast cells was detected by Western blotting. The yeast cells cotransformed with pAS2-1-X and normal human liver cDNA library were grown in selective SC/-trp-leu-his-ade medium. The second screen was performed with β-gal activity detection, and false positive clones were eliminated by segregation analysis, true positive clones were amplified,sequenced and analyzed with bioinformatics. Mating experiment was peformed to confirm the binding of putative proteins to X protein in the yeast cells. RESULTS: Bait plasmid pAS2-1-X was successfully constructed and pAS2-1-X correctly expressed BD-X fusion protein in yeast AH109. One hundred and three clones grew in the selective SC/-trp-leu-his-ade medium, and only one clone passed through 13-gal activity detection and segregation analysis. The inserted cDNA fragment showed high homology with Homo sapiens cytochrome C oxidase III (COXIII). Furthermore, mating experiment identified that the binding of COXIII to X protein was ***: COXIII protein is a novel protein that can interact with X protein in vivo by yeast two-hybrid system, and may contribute to the development of HCC through the interaction with X protein.
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