Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (12): 190-194.doi: 10.13560/j.cnki.biotech.bull.1985.2014.12.032

• Research Report • Previous Articles     Next Articles

Construction of Eukaryotic Expression Vector for the CD151 and CD163 Receptor Proteins of Porcine Reproductive and Respiratory Syndrome Virus

1,2Ruan Zheng,1Li Jie,2Liu Kaiwu,1Wang Lianfang,1Hua Juan,3Hu Xiaoming,1,3Liu Jie,1,4Huang Haijun   

  1. (1 .Wuhan Institute of Animal and Veterinary Science, Wuhan 430208; 2. Wuhan Animal Disease Prevention and Control Center, Wuhan 430023; Key Laboratory of Swine Breeding and Genetics, Ministry of Agriculture & Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction(Huazhong Agricultural University), Ministry of Education, Wuhan 430070; 4. Wuhan Bureau of Animal Husbandry and Veterinary, Wuhan 430023)
  • Received:2014-09-22 Online:2014-12-08 Published:2014-12-12

Abstract: In order to study the interaction mechanism between the PRRSV and cell receptors, the CD151 and CD163 code fragments were obtained from PAM cells by PCR. These PCR products were digested with Sal I and Kpn I, then inserted into pIRES2-EGFP vectors separately to construct eukaryotic expression plasmid pIRES2 EGFP - CD151 or pIRES2 EGFP - CD163. These vectors were identified by the methods of restriction enzyme digestion, PCR and sequencing. Marc 145 cells transfected with CD151 or CD163 or CD151/CD163 expressed strong green fluorescence, which was further confirmed by western blot. The successful establishment of eukaryotic expression vectors of CD151 and CD163 provides the material foundation for the further exploring the synergistic effect of CD151 and CD163 in the process of PRRSV infection, especially to the further study of PRRS virus invasion and the host identification.

Key words: Porcine reproductive and respiratory syndrome, CD151, CD163, Eukaryotic expression vector, Cell transfection