Isolation Of Porcine Reproductive And Respiratory Syndrome Virus (Prrsv) In Guangxi And Expression Of It's E Gene

Recently, some pig farms in Guang xi occur frequently an infectious disease,which characters are abortion,death of embryo,mummified fetuses,and respiratory problems in piglets . Pathogen of the disease was considered as porcine reproductive and respiratory syndrome virus (PRRSV). PRRS cause great lose and to control it is difficult without an effective vaccine. In this study, we collected samples from many farms in Guang xi and Guang dong.identified positive materials by RT-PCR using a pair of specific N gene primers of PRRSV. PAM and Marc-145 cells were inoculated with these positive samples. After two blind passages, typical cytopathogenic effect (CPE) was observed on cells as well as the positive control. And PRRSV was positive in the cells and culture medium by RT-PCR. The second PRRSV strain in our lab was isolated successfully.named GD strain.E gene was obtained from recombinant plasmids pMD-E by RT-PCR(PE1,PE2), then subcloned to prokaryotic expression vector PET-32a, PET-30a. And constructed the prokaryotic expression recombinant plasmids PET32a- E, PET30a-E.They were identified by being digested with restriction endonucleases , the results showed that these recombinant plasmids were succeessfully constructed .and then transport BL-21(DE3),induced with IPTG,we obtained gp5.i am sure we can make high quality vaccine on base of it in the near future.