Objective: To express recombinant tetanus toxin fragment C (rTTC) in prokaryotic cells, purify the expressed product and determine its activity.Methods: TTC gene fragment was amplified by PCR from the genomic DNA of Clostridium tetani and inserted into vector pThioHisA, and the constructed recombinant plasmid rTTC-pThioHisA was transformed to Escherichia coli BL21 for expression under induction of IPTG.The expressed rTTC protein was purified by ion exchange and gel filtration chromatog., and determined for activity and immunogenicity by Western blot, double immunodiffusion test and immunization of animals.Results: Restriction anal. showed that recombinant plasmid rTTC-pThioHisA was constructed correctly.The expressed rTTC protein existed in both soluble and inclusion body forms and reached a purity of more than 95% after purificationThe purified rTTC protein reacted with the antibody against complete tetanus toxin (TT), while the antibody against rTTC reacted with complete TT.The rTTC induced high immune response in rabbits but low immune response in mice.Conclusion: Recombinant TTC was expressed in prokaryotic cells and purified, which might be used as a novel vaccine against Clostridium tetani and a protein vector of bacterial polysaccharide vaccine.