HBV/HEV二价抗原的载体构建与表达

Construction and Identification of biological trait Assay of HBV/HEV Bivalent Antigen

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重组原核表达载体,采用PCR方法扩增出HBV的S基因,及HEV的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增强子的pT-T7高效的E.coli表达载体中。构建了pT-T7SE表达载体并且表达出相应的目的蛋白。该实验结果为研制预防乙型和戊型肝炎双价疫苗提供了实验技术基础。

In order to construct an high efficient expression vector of E.coli expressing bivalent vaccinal antigen which can resist HEV and HBV, S gene of The genome ofHBV adr subtype was amplified by PCR methods, furthermore, gene frangment encoding C-terminal 414- 606 aa of ORF2 region of HEV was amplified by PCR methods.Through T vector linkage and verdfication of colony PCR, double enzyme chopping, S gene and the partial gene frangment of ORF2 region were recombinated correctly into a high efficient E .coli expression vector pTβ-T7 containing Ω enhancer, as a result pTΩ-T7SE expression vector was constructed and the protein of interest could be expressed in E .coli. so that this result offered a good experiment basis for development the bivalent vaccine of the HEV and HBV hepatitis simultaneously.