The construction of prokaryotic expression plasmid and high-level expression of the human single chain Fv antibody fragment gene to rabies virus

To construct the prokaryotic expression plasmid of human scFv41 gene to rabies virus and prepare the scFv41 protein for identifying biological function. Using recombinant plasmid pCANTAB-scFv41 as template, the scFv41 gene with NcoI and NotI sites was amplified by PCR and cloned into the prokaryotic expression vector pET-22b(+). E.coli BL21(DE3) was transformed with the recombinant expression plasmid pET-scFv41 and protein expression was induced in Luria-Bertani medium by addition of IPTG. The recombinant expression plasmid pET-scFv41 successfully expressed scFv41 protein. The yield of the scFv41 fragment accounted for about 30% of the total bacterial protein and the protein showed specific immunological reaction with rabies virus in competition ELISA. The construction of the prokaryotic expression plasmid of scFv41 gene and preparation of scFv41 fragment established a solid basis for further studying the biological function 0f the human scFv41 to rabies virus.