人乙型肝炎病毒聚合酶在酿酒酵母中的表达  

Expression of Human Hepatitis B Virus Polymerase in Saccharomyces Cerevisiae

作　　者：于杨[1] 洪性出 

机构地区：[1]韩国全州市全北大学校医科大学微生物教研室

出　　处：《辽宁医学院学报》2008年第5期400-402,409,共4页Journal of Liaoning Medical University (LNMU) Bimonthly

摘　　要：目的尝试在酵母中表达人乙型肝炎聚合酶蛋白(HBV-Pol),并用镍基颗粒队蛋白粗提物进行部分纯化。方法将全长HBV-Pol基因用PCR方法扩增并连接到在酿酒酵母高效表达的质粒中。转化株于30℃培养3 d后进行诱导表达16 h,4℃下收集蛋白粗提物并用镍基颗粒进行提纯。用免疫印迹技术检测表达结果。结果重组的HBV-Pol存在于该型酵母的蛋白粗提物中,该蛋白产物可被Nickel-particle提取。结论本研究为实现HBV-Pol在S.c.中的高水平表达提供了可靠依据,也便于HBV-Pol的生化特性研究以及蛋白纯化和抗体制备的进一步开展。Objective Attempt to express HBV - Pol in yeast expression system and then partially purify the rough extract with nickel - based particle. Methods Full length HBV - Pol gene was amplified and was introduced into a high effective expression plasmid of Saccharomyces cerevisiae （ S. c. ）. The transformant was cultured at 30℃ for 3 days, and then induction was carried out for another 16 hours at 30℃, and then induction was carried out for another 16 hours at 30℃. Cell lysis and purification were preformed at 4℃. Target protein was detected in harvested rough extract with western blot. Results The recombinant HBV - Pol was present in rough extract of yeast and could be purified with nickel particle. Conclusions Results of this study prove that HBV - Pol can be expressed in S. c. yeast and might facilitate the biochemical study, protein purification and antibody preparation of HBV - Pol.

关 键 词：乙型肝炎病毒 聚合酶 酿酒酵母 

分 类 号：R512.62[医药卫生—内科学]