ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2012, Vol. 43 ›› Issue (8): 1266-1272.doi:

• 预防兽医 • Previous Articles     Next Articles

Construction and Humoral Immune Response of Recombinant Plasmid CoExpressing GP3,GP5 and M Proteins of Porcine Reproductive and Respiratory Syndrome Virus

WANG Fan 1, LIU Jianbin 2, ZHU Xiumei 1, LV Zhihui 1, MA Quanying 1,

DU Ping 1, LIU XueRong 1, MU Kebin 1*

  

  1. 1. China Agricultural Veterinarian Biology Science and Technology Co., Ltd, Lanzhou 730046, China； 2. Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China
  • Received:2012-01-11 Revised:1900-01-01 Online:2012-08-24 Published:2012-08-24
  • Contact: MU Kebin

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Abstract: The aim of this study was to construct a recombinant eukaryotic plasmid coexpressing GP3, GP5 and M proteins of porcine reproductive and respiratory syndrome virus (PRRSV). GP3, GP5 and M protein gene fragments of PRRSV LN strain were amplified by PCR and cloned into vector pcDNA3.1(+) to produce the recombinant plasmid pcDNA3.1GP3GP5M. The plasmid was transfected into COS7 cells, and immunological responses to the plasmid were investigated in BALB/c mice and piglets. The recombinant plasmid pcDNA3.1GP3GP5M was transfected into COS7 cells, PCR identification and indirect immunofluorescence assay proved that GP3, GP5M gene of PRRSV were expressed in COS7 cells. The result of Western blotting showed that the GP3 and GP5 and M proteins were coexpressed and formed fusion protein. The BALB/c mice were injected with recombinant plasmid pcDNA3.1GP3GP5M to evaluate the induced immunological responses in vivo. The specific detectable antiPRRSV neutralizing antibodies were produced in the vaccinated mice at 2 weeks and reached a peak 1:32 at 8 weeks after primary vaccination. In addition, it was observed that the 1:41:8 neutralizing antibodies of the vaccinated piglets. The results showed that the recombinant plasmid pcDNA3.1GP3GP5M has been constructed successfully, and the recombinant plasmid has well immunity, the recombinant plasmid could be a basis to eukaryotic vector vaccine for PRRSV infection.