title: [Construction and expression of trichosanthin mutant gene TCS(RL28-29CG) and purification of expressed product].
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  TCS mutant gene TCS(RL28-29CG); is succ-essfully constructed and expressed.
authors.affiliation: Department of Microbiology, The Fourth Military Medical University, Xi'an 710032, China. judy_625@163.com
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Shi
authors.forename: Zi-yu
authors.initials: ZY
authors.lastname: Liu
authors.forename: Zhi-kai
authors.initials: ZK
authors.lastname: Xu
authors.forename: Jing
authors.initials: J
authors.lastname: Xu
authors.forename: Zhi-dong
authors.initials: ZD
authors.lastname: Li
mesh_terms: D015153:Blotting, Western; D004926:Escherichia coli / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000302:isolation & purification*; D029778:Trichosanthes / Q000235:genetics*; D015978:Trichosanthin / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D050505:Mutant Proteins; D015978:Trichosanthin