Objective:To construct and express a recombinant immunotoxin expression vector composed of a human anti-c-Met single-chain Fv fragment gene and a PE38KDEL gene,and to examine the binding ability of the purified product to human hepatic cellular cancer cell line SMMC 7721. Methods:The human anti-c-Met gene fragments were amplified through PCR then it was subcloned into a PE38KDEL gene inserted fusion protein expression plasmid,which was expressed by pBAD/GⅢA vector. The recombinant vector was identified by restriction endonucleases digestion,PCR and DNA sequencing. After transformed into E.coli Top10F′and induced by L-arabinos,the inclusion bodies were denatured,renatured and refolded,then resolvable protein was purified. The binding ability and effect on cell proliferation test of the purified protein was evaluated by FACS and MTT,respectively. Results:The expression vector pBAD/GⅢA/c-Met/PE38KDEL was constructed successfully and the expression product existed mainly in the form of inclusion body. The refolding product remained the binding ability of the single-chain antibody and had significant effect of cell toxicity. Conclusion:Results of our study on c-Met/PE38KDEL and its effects on SMMC7721 show that the fusion protein would be a promising candidate for tumor targeted immunotherapy.