马动脉炎病毒大囊膜糖蛋白真核表达载体的构建与瞬时表达

Construction and transient expression of equine arteritis virus major envelope glycoprotein of eukaryotic expression vector

将马动脉炎病毒大囊膜糖蛋白基因GL插入真核表达载体pVAX1中构建真核表达载体pVAX1-GL,酶切和测序结果表明构建是正确的,用脂质体转染试剂将其转染BHK-21细胞并通过间接免疫荧光试验检测其在体外的表达情况,结果在转染的细胞表面观察到绿色荧光,证明基因得到了表达,而对照则无绿色荧光,本研究为马动脉炎基因疫苗的研究奠定了基础。

To construct the eukaryotic expression vector of equine arteritis virus major envelope protein gene and observe its expression in vitro, the recombinant expression vector pVAX1-GL was constructed by inserting the GL gene into the eukaryotic expression vector pVAX1. The pVAX1-GL was transfected into BHK-21 cells by lipofectamine and the expressed product was detected by indirect immunofluorescence. Restriction enzymes digestion analysis and sequencing results revealed that the recombinant expression vector pVAX1-GL has been constructed successfully. The indirect immunofluorescence result showed green fluorescence on the membrane of transfected cells. The constructed eukaryotic expression vector of EAV-GL can be expressed in vitro, which lay the foundation for the further study of EAV DNA vaccine.