Expression Vector Construction And Its Immunogenicity Of The Recombinant Adenovirus E Protein Of West Nile Virus To Mice

West Nile virus can cause serious disease in human and animal.Human and animal infected with WNV, which emerge such as fever,meningitis and encephalitis,even resulting death.Therefore WNV received great attention worldwide.At present, there are no report of clinical case caused by WNF in China. However, complex geographical environment of China and many kind of meda such as mosquito will result in fast propagation, study on WNV is more significant in entry and exit supervision and prevention system in China. WNV and JEV belong to the same family, and there are immunologic cross-reactivity and serological cross-reaction. In China, JEV has been confirmed that have been existing for a long time, but not exist clinical symptoms of WNV. So to explore the immune cross reaction and serological cross reaction cross immune protection against virus belong to the same family, as well as cross-immunity protection function of split vaccine development has provided the possibility.E protein is the most important structural protein in surface of WNV, which epitope is relate to neutralization activity, tissue tropism, serological specificity, Virulence,host range. E protein polypeptide chain folds formed through three institutions Domain (Domain Ⅰ、Ⅱ and Ⅲ). DomainⅢ is the binding receptor binding domain, which also is B cell epitope of neutralizing antibodies. Neutralizing antibody combine with E protein can blocked WNV combined with corresponding receptors on cells, or blocked viruses and cell membrane fusion.In this study, WNV E protein was expressed through prokaryotic expression vector to expression and identified by Elisa and WB, and was used it to make positive serum to E protein. Then, a recombinant eukaryotic expression vector was constructed and transfected into BHK-21cell, identified by IFA to find E protein expressed in cells.Positive serum can be used for further experiments.To construct recombinant adeno virus expressing E protein of WNV, the WNV E gene was cloned into the shuttle plasmid vector pshuttle-CMV. TThe recombinant virus of rAd-WNV-E was generated by transfecting the Pac I linearized pAd-WNV-E into Ad-293cells. The WNV E protein was efficiently expressed from the recombinant virus detected by western blot and immunofluorescence assay, and the viral titration results showed that the replication ability of rAd-WNV-E was the similar to the wild type recombinant virus. These results provided the basis for further study on the recombinant virus vaccine against WNV.In animal experiments, rAd-WNV-E in mice can produced E protein antibody titer and has high specificity, but cross-protection among infection of JEV mice immune protection is very weak, relative only survival percent15%.