Expression of GP5-M fusion protein of porcine reproductive and respiratory syndrone virus (PRRSV) and establishment of ELISA diagnose based on the recombinant fusion protein.
- Author:
Yun-Bo JIANG
1
;
Liu-Rong FANG
;
Shao-Bo XIAO
;
Tian-Tian XIE
;
Huan-Chun CHEN
Author Information
1. Laboratory of Animal Virology, College of Animal Science and Veterinary Medicine, Huazhong Agriculture University, Wuhan 430070, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Enzyme-Linked Immunosorbent Assay;
Escherichia coli;
genetics;
metabolism;
Glutathione Transferase;
metabolism;
Open Reading Frames;
Porcine Reproductive and Respiratory Syndrome;
diagnosis;
immunology;
Porcine respiratory and reproductive syndrome virus;
genetics;
immunology;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Swine;
Viral Envelope Proteins;
biosynthesis;
genetics
- From:
Chinese Journal of Biotechnology
2005;21(2):259-264
- CountryChina
- Language:Chinese
-
Abstract:
The cDNA fragment encoding the truncated GP5 and the full-length M protein of Porcine Reproductive and Respiratory Syndrone Virus (PRRSV) were orderly fused to the downstream of glutathione S-transferase (GST) of pGEX-KG expression vector, resulting in the fusion expression plasmid pKG-56. After transformed into E. coli BL21 (DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-GP5-M fusion protein was expressed in high level. Western-blot was performed to confirm that the expressed fusion protein could specifically react with antiserum against PRRSV. The fusion protein was further purified and used as an antigen to establish a novel PRRSV ELISA diagnose assay (P56-ELISA). Comparison between P56-ELISA and the abroad kit IDEXX-ELISA showed the two methods had 94.1 percent agreement by detecting 205 serum samples, indicating that the indirect P56-ELISA was specific and sensitive. The correlation between virus neutralization antibody of the infected pigs (not convalescent pigs) and antibody response to the fusion protein GP5-M was further studied. The regression function analysis suggested that there was no significant correlation between ELISA antibody response (OD630 nm) to the fusion protein GP5-M in clinical serum and their specific neutralizing titers.
