Expression,optimization and identification of tetanus toxin fragment C in E. coli BL21(DE3)

TTc was obtained by PCR through base mutant upstream primer.After a series of DNA recombination manipulation,expression plasmid pET-42(b)/TTc was finally constructed and transformed into E.coli BL21(DE3) for IPTG-induced expression.Expressed protein is soluble,the rate of target protein is about 22%,and MW is about 50kD.After optimization of medium,inducing time,inducing temperature,solubility and expression level of target protein are all improved.Western blotting assay showed that expressed can react with TTc McAb.This study result provide the basis for developing this protein as a protein carrier and recombinant vaccine.