Mahnke C, Löchelt M, Bannert H, Flügel R M
Institut für Virusforschung, Deutsches Krebsforschungszentrum, Heidelberg, F.R.G.
J Virol Methods. 1990 Jul;29(1):13-22. doi: 10.1016/0166-0934(90)90003-x.
Recombinant plasmid clones were constructed harbouring the central domains of the outer membrane protein and the transmembrane protein of the env gene of human spumaretrovirus (HSRV). The corresponding fusion proteins were expressed in E. coli, purified and used subsequently to produce antibodies against the HSRV env proteins in rabbits. The authenticity of the bacterially produced domain of the HSRV env proteins was shown by radioimmunoprecipitation of the viral env glycoprotein from HSRV-infected human cells with rabbit antibodies raised against the recombinant antigens. The recombinant viral antigens were used to establish a sensitive and spumavirus-specific enzyme-linked immunosorbent assay (ELISA). This anti HSRV antibody ELISA makes it possible to screen human sera for the presence of spumavirus infections.
构建了重组质粒克隆,其含有人类泡沫逆转录病毒(HSRV)包膜基因的外膜蛋白和跨膜蛋白的中央结构域。相应的融合蛋白在大肠杆菌中表达、纯化,随后用于在兔体内产生针对HSRV包膜蛋白的抗体。通过用针对重组抗原产生的兔抗体对HSRV感染的人类细胞中的病毒包膜糖蛋白进行放射免疫沉淀,证明了细菌产生的HSRV包膜蛋白结构域的真实性。重组病毒抗原用于建立一种灵敏的、泡沫病毒特异性的酶联免疫吸附测定(ELISA)。这种抗HSRV抗体ELISA能够筛查人血清中是否存在泡沫病毒感染。