Construction And Immunogenicity Evaluation Of Recombinant Adenovirus Expressing VP1of Porcine Teschovirus-8

The porcine teschoviruse (PTV) infected pigs was described as causative agents of severe andmild neurological disorders, reproductive failure, pneumonia, diarrhea, pericarditis, myocarditis anddermal lesions of swine. In recent years，PTV was frequently accompanied by porcine reproductive andrespiratory syndrome virus (PRRSV) and porcine circovirus (PCV2) to co-infect pigs, which hamperedthe identification, diagnosis of the disease and caused economic losses in the pig industry. In order toprevent the PTV infection and prepare for the vaccine, the recombinant adenovirus containing VP1gene of porcine teschovirus-8was constructed and the immunogenicity of the recombinant adenoviruswas evaluated in this study.To develop a recombinant adenovirus vector containing VP1gene of porcine teschovirus-8, a pairof primers targeting on VP1gene of porcine teschovirus-8were designed and then VP1gene wereamplified and cloned into tansfer vector according to the manufacturer’s instruction. The recombinanttansfer vector was identified by PCR and enzyme digestion, then linearized by PmeⅠafter sequencedand co-transformed into Escherichia coli strain BJ5183containing pAdEasy-1vector by electroporationto achieve homologous recombination. The recombinant adenovirus vector was identified by PCR andPacⅠdigestion. Then the plasmid was linearized by PacⅠand transfected into293cells to formrecombinant adenovirus. RT-PCR, indirect immunofluorescence assay (IFA) and Western blot wereused to identify the expression of VP1. The immunogenicity of recombinant adenoviruses wasexamined in3-week piglet not infected with PTV by vaccination with the recombinant adenovirustwice at three-weeks intervals. The results showed that the recombinant adenovirus could providehumoral immunity and cellular immunity responses in pigs by the detection of virus neutralizationtest(VNT), indirect ELISA and lymphocyte proliferation response.