Expression Of The Surface Protein Of JSRV Env Gene And Preparation Of Monoclonal Antibodies

Sheep pulmonary adenomatosis (SPA),a contagious lung neoplasia of sheep is caused by the Jaagsiekte Sheep Retrovirus(JSRV),a betaretrovirus. According to the OIE classification, SPA belongs to the B-type infectious disease.The aim of this study was to investigate the pathogenesis of JSRV and diagnosis method of SPA. In this study the recombinant plasmids pGEX-4T-1-SU we saved was transformed to E.coli BL21. After expression and purification , it was used as immunogen to immunize six-week-old to eight-week-old BALB/c mice. Three hybridoma cell lines named 1D6,3B3,3D6 ,were obtained by using hybridoma technique, respectivly. All the cell lines being screened by indirect ELISA, in which the pured recombinant protein pGEX-4T-1-SU and the protein GST what was expressed by the expression vector pGEX-4T-1 were both used as antigen.To obtain monoclonal antibodies against the natural protein, the SU gene was cloned into a plasmid pCDNA3.1(+), containing a cytomegalovirus promoter. When the recombinant plasmid SU-HA-pcDNA3.1(+) was transfected into A-549 cells, mimic natural protein was expressed. This protein was used as antigen to detecet the three hybridoma cell lines by indirect immunofluorescence assay experiments(IFA).The results showed that the three were all positive. It was said that three hybridomas cells which can product McAb against SU of JSRV-NM strain steadily had been established. The results reported above laid certain foundation for explaining the pathogenesis of SPA, for establishing a sensitive diagnosis method of JSRV, for analying the function of JSRV env gene surface protein, etc. Consequently, it has very important academic and practical significance for ensuring the security of study sheep industry and the health development of sheep-rearing.