应用杆状病毒载体表达狂犬病病毒糖蛋白

Expression of Glycoprotein of Rabies Virus Using Baculovirus Vector

本试验利用杆状病毒载体成功地表达狂犬病病毒糖 (G) 蛋白。从转染重组了日本动物用狂犬病病毒疫苗株RC HL株G蛋白基因的转移载体和野生型杆状病毒DNA 的Sf 9 细胞中, 分离出2 个重组杆状病毒克隆。应用印渍法和荧光抗体法(IFA) 从重组杆状病毒感染的Sf 9 细胞检出了狂犬病毒G 蛋白。Sf9 细胞表达的G 蛋白与狂犬病毒RC HL株感染NA 细胞的G蛋白的分子量一致。35 个抗RC HL株G蛋白的单克隆抗体均与重组杆状病毒感染的Sf 9 细胞反应, 表明表达的G蛋白的抗原性没有发生变异。表达的G蛋白主要分布在Sf 9 细胞的膜表面, 形成类似环状的荧光,这种荧光与RC HL株感染的NA 细胞的荧光相同。

In this experiment we have successfully expressed rabies vivus glycoprotein(G protein) using baculovirus vector.Two clones of recombinant baculovirus were isolated from Sf9 cells,in which was transfeted with DNA of wildtype baculovirus and baculovirus transfer vector inserted G protein gene of RCHL strain used for production of animals rabies vaccine in Japan.By using western blot and indirect immunofluorescent assay(IFA),the G protein of rabies virus was detected from Sf9 cells which were infected with recombinant baculovirus.The G protein in Sf9 cells expressed by recombinant baculovirus showed identical molecular weight with that in NA cells infected with RCHL strain of rabies vrus.Thirtyfive monoclonal antibodies(MAbs) against G protein of RCHL strain reacted with G protein on the Sf9 cells infected with recombinant baculovirus,indicating that antigenic variation of the expressed G protein has not been found.Majority of the expressed G protein exists on the surface of Sf9 cells,fluorescence of the Sf9 cells formed a ringlike when it infected with recombinant baculovirus and was dyed by IFA.This kind of fluorescence on the Sf9 cells was the same as that on NA cells infected with RCHL strain of rabies virus using IFA.