hDaxx与乙型肝炎病毒X蛋白的相互作用  

Interaction of hDaxx and Hepatitis B Virus X Protein

作　　者：周艺[1] 李金丽[2] 万艳平[2] 

机构地区：[1]南华大学公共卫生学院,湖南衡阳421001 [2]南华大学病原生物学研究所

出　　处：《南华大学学报（医学版）》2008年第3期279-281,共3页Journal of Nanhua University(Medical Edition)

基　　金：湖南省自然科学基金课题(NO:06JJ4108)

摘　　要：目的采用酵母双杂交法研究HBVX蛋白与hDaxx蛋白的相互作用，为探讨HBVX蛋白的致癌机制奠定实验基础。方法构建pGBKT7-hDaxx和pGADT7-HBX重组质粒，分四组转化酵母AHl09，A组为pGADT7和pGBKT7，B组为pGBKT7-hDaxx和pGADT7-HBX，C组为pGADT7-T和pG-BKT7-Lam，D组为pGADT7-T和pGBKT7-p53。将转化菌落接种于SD／-Tip-ku（二缺）固体平板，长出克隆后再接种于SD／-Tip-Leu-His（三缺）和SD／-Tip-Leu-His-Ade（四缺）平板，30℃培养36～72h。裂解酵母菌，提取蛋白，经SDS-PAGE、Western blot检测hDaxx和X蛋白在酵母中的表达。结果仅有共转化pGBKT7-hDaxx和pGADT7-HBx的AHl09可以在三缺及四缺平板上生长。Western blot检测到hDaxx和X蛋白均能在酵母中表达。结论HBV X蛋白与hDaxx能在酵母细胞内发生相互作用。Objective To identify the interaction of Hepatitis B virus X protein and hDaxx by yeast two-hy-brid system to explore the contribution of HBx protein to hepatocellular carcinoma. Methods The vector of pGBKT7-hDaxx and pGADT7-HBx were constructed for yeast two-hybrid system. Then four groups of different plasmids were divided for transforming into yeast （AH109）, respectively. Group A was pGBKT7-hDaxx; group B was pGADT7-HBx; group C was pGBKT7 and pGADT7 ; group D was pGBKT7 - hDaxx and pGADT7-HBx. The transformants were plated on SD/Trp-Leu plates and incubated at 30℃ for 48h. The positive clones were streaked to be assayed on SD/Trp-Leu-His plates and SD/Trp-Leu-His-Ade plates, and then incubated at 30℃ for 48 - 96h. The expression of hDaxx and X protein were detected by Western blot. Results The AH109 which were eotransformed the plasmids of pGBKT7-hDaxx and pGADT7-HBx grew on SD/Trp-Leu-His plates and SD/Trp-Leu-His-Ade plates. The expression of hDaxx and HBx protein were demonstrated by Western blot. Conclusion HBV X protein could interact with hDaxx in yeast two-hybrid system.

关 键 词：HDAXX 乙型肝炎病毒X蛋白 酵母双杂交 

分 类 号：R373.21[医药卫生—病原生物学]