乙型肝炎病毒C基因片段的PCR扩增和克隆及其在大肠杆菌中的高效表达

PCR AMPLIFICATION,CLONING AND HIGH LEVELEXPRESSION IN E. COLI OF HEPAIITIS B VIRUS C GENE

乙型肝炎病毒Ｃ基因片段的ＰＣＲ扩增和克隆及其在大肠杆菌中的高效表达芦睿，张楚瑜，李小峰，丁建华（武汉大学生命科学院病毒学系武汉４３００７２）关键词ＨＢｃＡｇ，ＨＢｅＡｇ，ＰＣＲ，基因表达根据目前对ＨＢｃＡＧ和ＨＢｅＡｇ及其相互关系的认识，我们通过聚合...

new recombinant expression plasmid pHBc149 was constructed by inserting the DNAsegment of 447 base pairs from the 5’-terminal of HBcAg gene,obtained by polymerase chain reaction,into expression vector pBV220 containing PRPL promoter and cIts857temperature sensitive gene. After transforming to E.coli,we obtained a high level expressionstrain.Shifting the culture temperature from 30℃ to 42℃ led to a high level expression of the recombinant polypeptide,which could self -assemble into particles.SDS-PAGE analysis showed that the expressed product of pHBc 149 accounted for 30.41 percent of all theproteins,with a molecular weight of about 16KD.The ELISA titer of the pHBc149 product was1∶320,000 to HBcAg and 1∶160,000 to HBeAg.Further treated with denaturant ,the pHBc149product could be transformed into HBeAg with novel bioactivity.