Construction and Verification of Anti-Mm Scfv-Tp Fusion Protein Expression Vector

Objective To construct an expression vector of anti-MM scFv-tP fusion protein and test its expression efficiency and function.Methods The truncated protamine (tP) gene sequence was added to the gene of single chain antibody against the specific antigen on the surface of malignant melanoma tumor cells using PCR.A GST-fusion expression vector was constructed and the soluable protein was expressed in the E.coli system.After cleavage and purification,the purified fusion protein was obtained.The binding activity of Anti-MM scFv-tP and siRNA was detected by EMSA.Flow cytometry and confocal microscopy were used to detect the cell surface antigen binding activity of the fusion protein.Results The expression vector of Anti-MM scFv-tP fusion protein was successfully constructed.The soluable protein could be expressed in the E.coli system,and the purified fusion protein was obtained.The anti-MM scFv-tP fusion protein retained siRNA binding ability and could directly target malignant melanoma (MM) LiBr cells.Conclusion The recombinant GST-Anti-MM-scFv-tp expression vector was successfully constructed.The fusion protein retains siRNA binding ability and can directly target LiBr cells to provide a reliable tool for further study.