Construction and Immunogenecity of GP5 Recombinant Adenovirus of Porcine Reproductive and Respira- tory Syndrome virus

The GP5 gene of Porcine reproductive and respiratory syndrome virus (PRRSV) S1 strain was amplified by RT-PCR, which was digested with KpnI and XhoI and cloned into pShuttle-CMV plasmid. Sequence analysis showed that GP5 gene was cloned in correctly. Then the recombinant plasmid pShuttle-CMV-GP5 was linearized , and co-transformed into E.coli BJ5183 strain together with adenovirus backbone vector pAdEasy-1. Recombinant adenovirus plasmid pAd-GP5 was linearized with PacI, and transferred into HEK293-A cells to obtain recombinant adenovirus. The obtained recombinant adenovirus rAd-GP5 was identified with RT-PCR and IFA. The result showed that the recombinant adenovirus expressed GP5 protein of PRRSV successfully. Virus neutralization assay was carried out after callection of serum, the result demonstrated that the recombinant adenovirus can elicit effective neutralization antibody against of GP5 PRRSV, therefore, the recombinant adenovirus may be a candidate for gene engineering vaccines.