Prokaryotic Expression of PRRSV GP2-GP4 Protein and Preparation of Polyclonal Antibody
Porcine reproductive and respiratory syndrome virus(PRRSV)is one of the important pathogens that harm the development of pig industry.Since the structural proteins GP2 and GP4 can bind to the CD163 receptor of target cells and can participate in the virus neutralization reaction,the study for the GP2 and GP4 protein is significant for PRRSV vaccine development.In this study,GP2 and GP4 genes were linked by flexible peptides,cloned into the prokaryotic expression vector pET32a,and the GP2-GP4 fusion protein was successfully expressed through the expression system of E.coli.The target protein was expressed in the form of an inclusion bodies,and high purity GP2-GP4 fusion protein was obtained after purification and refolding.The polyclonal antibodies were prepared by immunizing rabbits.SDS-PAGE results showed that the size of the target protein was about 55 kDa,which was consistent with the expectation.ELISA revealed a GP2-GP4 antibody titer of 1∶64000.Western blot and IFA results showed that polyclonal antibody could react specifically with GP2-GP4 protein with good reactivity.In summary,the fusion expression of GP2-GP4 proteins was obtained and the polyclonal antibody was prepared with good specificity,which laid a foundation for the investigation of the function of PRRSV GP2-GP4 proteins and subsequent vaccine research.
porcine reproductive and respiratory syndromeglycoproteinprokaryotic expression
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