酵母双杂交文库筛选HBX相互作用蛋白

The screening of HBX interacting proteins by yeast two-hybrid library

目的:筛选鉴定新的乙肝病毒X蛋白(hepatitis B viral X protein,HBX)相互作用蛋白。方法:以HBX为诱饵蛋白,将其与酿酒酵母转录因子GAL4 DBD(DNA binding domain)融合表达,将人肝脏cDNA文库编码的蛋白与GAL4的AD区(activation domain)融合表达。通过酵母双杂交筛选鉴定肝脏中与HBX相互作用的蛋白。筛选得到的阳性酵母转化子,经质粒提取和测序后,分析其所包含的序列编码的蛋白,通过再次验证,初步确定该蛋白是否与HBX具有相互作用。结果:筛选得到5个潜在的能与HBX相互作用的蛋白。结论:通过酵母双杂交c DNA文库筛选,得到多个潜在的HBX相互作用蛋白,进一步的验证及功能研究将对揭示HBX促进肝癌发生的机制起到重要的推动作用。

Objective：To Screen and identify new HBX （Hepatitis B viral X protein） interacting proteins. Methods： HBX was used as a bait protein, and it was fused with the Saccharomyces cerevisiae transcription factor GAL4 DBD （DNA binding domain）, while the protein encoded by the human liver cDNA library was fused with the AD domain of the GAL4 （Activation domain）. Proteins interacting with HBX in the liver were identified by yeast two-hybrid library screening. The positive yeast transformants were selected. After plasmid extraction and sequencing, the proteins encoded by the sequences were analyzed. Finally, the proteins potentially interacted with HBX were reconfirmed by yeast two-hybird. Results： Through yeast two-hybrid library screening, 5 potential proteins that could interact with HBX were identified. Conclusion： Five potential HBX interacting proteins were obtained by yeast two-hybrid cDNA library screening. Further validation and functional studies will play an important role in illuminating the mechanism by which HBX promotes hepatoma carcinoma.