The Rubella Virus E1 Fusion Of Prokaryotic Expression Clone Construction And Expression

Construction and Expression of the Prokaryotic Expressive Clone of Rubella Virus E1 Fusion ProteinPostgraduate student : Lu Ning Tutor : Prof. Wang BinDepartment of Molecular Virology,Medical College of Qingdao UniversityAbstractObjective To construct the rubella virus (RV) E1 antigen procaryotic expressive clone with the aim of developing efficient recombinant antigens for serological assays of RV infection. Methods RT-PCR was employed to amplify the target gene (nt8886-8976) coding for RV E1 212 - 242 amino acid residues, the main antigenic determinants of RV. The amplifed fragments were then cloned to the procaryotic expressive vector pGEX4T-2 and transfomated to E. coli. DH5α. The E1 recombinant clone was induced with IPTG to express the E1 fusion proteins and the products were purified by affinity chromatography. The antigenecity of the proteins were tested by Western-blot . Results E1 fusion proteins (35KD) were efficiently expressed in E. coli. DH5α. In Western-blot assay, the fusion proteins identified 8 of 11 RV IgM-positive serum samples(72.7%) and reacted with neither of the 5 negative serum samples. Conclusion The RV E1 fusion proteins remaining good antigenicity were successfully expressed in E. coli., which has provided useful information for development of RV recombinant antigens.