Expression and purification of mutant trichosanthin in escherichia coli

Objective: To construct a mutated trichosanthin(mTCS) in which four anino acids(Aa120-123) are deleted and to express and purify this mTCS in E.coli.Methods: cDNA encoding mTCS was obtained by PCR-driven overlap extension,and then cloned into pET-28a(+) vector.After restriction and sequence analysis,the recombinant plasmid pET-28a(+)-mTCS was transformed into BL21(DE3).The mTCS was expressed by IPTG induction and purified by Ni-NTA resin.Results: mTCS cDNA was obtained and cloned into the pET-28a(+) vector successfully.In BL21(DE3),the mTCS was expressed by IPTG induction,and purified by Ni-NTA resin efficiently.Conclusion: A mutated TCS was obtained in this study and a technique method for prokaryotic expression and purifaication of this mutated TCS was established successfully.