【摘要】 [目的]为构建一个能够表达出乙/戊型肝炎二价疫苗抗原的大肠杆菌高效表达载体。[方法]从乙型肝炎患者血清中提取adr型基因组,采用PCR方法扩增出S基因;从戊型肝炎患者的粪便中提取RNA,采用RT-PCR的方法扩增出ORF2编码区的羧基末端中的414-606aa的基因片段,经过T载体连接;菌落PCR;双酶切;测序鉴定后,将这两部分重组入含有Ω增强子的pTΩ-T7高效的E.coli表达载体中。[结果]构建了pTΩ-T7SE表达载体并且表达出相应的目的蛋白。[结论]该方法为同时预防乙型和戊型肝炎提供了一个较好的途径。更多还原

【Abstract】 [Objective]To construct an high efficient expression vector of E.coli expressing bivalent vaccinal antigen which can resist HEV and HBV.[Methods]The genome of adr subtype was extracted from the serum of B type hepatitis patients and S gene was amplified by PCR method.The RNA of HEV was extracted from manure of E hepatitis patients and gene frangment encoding C-terminal 414-606 aa of ORF2 region was amplified by RT-PCR method.Through T vector linkage and vertification of colony PCR,double enzyme chopping and sequencing,S gene and the partial gene frangment of ORF2 region were recombinated correctly into a high efficient E.coli expression vector pTΩ-T7 containing Ω enhancer.[Results]pTΩ-T7SE expression vector was constructed and the protein of interest was expressed in E.coli.[Conclusions]This measure offers a good approach to the prevention of HEV and HBV hepatitis simultaneously.更多还原