The Construction Of WSSV Gene Expression Library And Localization Of Ubiquitin Ligase Enzyme WSV249

White spot syndrome virus (WSSV) is one of the major pathogens of shrimp and other crustaceans. According to genome sequencing, WSSV contains more than180open reading frames. Although some proteins have been identified, the functions of a large number of proteins remain unknown.For the purpose of WSSV proteomics research, we used iel promoter core region nt(-94+52) and T7promoter to construct a shuttle vector that is capable of expressing WSSV proteins in both eukaryotic cell and prokaryotic cell. In addition, we successfully use the shuttle vector to clone162WSSV genes. Among these clones,94proteins can be expressed in E.Coli and102proteins can be expressed in High-Five insect cell. Our work lays a solid foundation for the future research about the function of WSSV proteins.WSV249has been proved as an E3ubiquitin ligase in the ubiquitin-proteasome pathway, but its substrate has not been found yet. In previous study, we found WSV249expressed around the nucleus. To identify the accurate location of WSV249, we expressed WSV249in both High-Five cell and293T cell. We confirmed that WSV249was located in the centrosome according to the result of immunofluorescence colocalisation. This result will contribute to future identification of the substrate of WSV249.