Abstract:ABSTRACT:The purpose of this study is to construct fusion proteins that can generate agglutination and immune reaction. Specific primers were used to amplify, from the recombinant plasmid pMD-2E8ScFv and pMD-G and with PCR, 2E8 gene (ScFv gene against H antigen of human erythrocytes) and Kg gene (the major antigen epitope domain of G gene), after which the two were spliced into a fusion gene by splicing overlap extension PCR method to construct prokaryotic expression plasmid pET-Trx-2E8Kg, and was then transformed into competent cell BL21(DE3)plysS. Finally, protein expression was induced with IPTG and analysed with SDS-PAGE.The results showed that the target fusion protein was successfully expressed and identified in inclusion bodies, with an expression accounting for 23.5% of the total bacterial protein, the molecular weight was 77.7 kD, consistent with the expected size. The purity of the expressed protein reached 98.9% after being purified by affinity chromatography and refolded by Glutathione reduction. The results of Western-blot detection and agglutination test on red blood cell indicated that the fusion protein 2E8kg had bifunctional characteristics, not only being able to have specific reaction with anti-rabies hyperimmune serum, but could combine with human red blood cell.