Aim: To construct and express a trichosanthin(TCS)gene mutant and purify the expressed product.
Methods: Predict the potential antigenic determinant on TCS molecule by computer modeling and induce site-directed mutation. Amplify gene mutant TCS(RL28-29CG); by PCR using the genomic DNA of Trichosanthes kirilowii as a template and insert into expression vector pRSET-A, then transform to E.coli BL21(DE3)for expression under induction of IPTG. Purify the expressed product by Ni-NTA afinity column chromatography.
Results: The target protein in a soluble form was successfully expressed in E.coli. Homogenous TCS mutant protein was obtained after purification of expressed product.
Conclusion: TCS mutant gene TCS(RL28-29CG); is succ-essfully constructed and expressed.