H3N8亚型EIV HA基因在昆虫细胞中的表达及生物学特性分析

Expression and biological characterization of HA protein of H3N8 subtype equine influenza virus in insect cells

为表达H3N8亚型马流感病毒(EIV)HA蛋白,本研究将A/equine/xinjiang/3/2007(H3N8)的HA基因克隆至杆状病毒转移载体pFastBac1中,构建重组转移质粒pFB-XJ3-HA,并将其转化至DH10BAC感受态细胞中,与杆状病毒骨架质粒Bacmid进行重组,重组杆粒rBac-XJ3-HA转染Sf9昆虫细胞,构建重组杆状病毒rBv-XJ3-HA,并通过细胞免疫组化和western blot进行鉴定。结果表明rBv-XJ3-HA构建正确,表达的HA蛋白与抗EIV的血清反应良好;血凝试验结果表明表达的HA蛋白具有良好的血凝活性。本研究为研制EIV HA蛋白亚单位疫苗提供了实验依据。

To express the hemagglutinin （HA） protein of H3N8 subtype equine influenza virus （EIV）, the HA gene of E1Vs A/equine/xinjiang/3/2007 （H3N8） was sub-cloned into transfer plasmid pFastBacl and then the recombinant shuttle plasmid pFB-XJ3-HA was transformed into DH10BA competent cells to generate a recombinant baculovirus bacmid of rBac-XJ3-HA. The rBac-XJ3-HA was transfected into Sf9 insect cells for packing a recombinant baculovirus of rBv-XJ3-HA which was confirmed by PCR, immunohistochemistry, western blot, and hemagglutination test. The results showed that HA protein of E1V was expressed in insect cells and have the immunoreactivity and hemagglutinin activity, which provide the basis for the development of subunit vaccine of H3N8 subtype equine influenza.