title: [Expression and analysis of recombinant chicken IL-18 in Pichia pastoris.].
abstract: OBJECTIVE
  Expression and analysis of recombinant chicken IL-18 in Pichia pastoris.
  
  
  METHODS
  Chicken IL-18 mature peptide gene was amplified from the recombinant plasmid pMD18-T-ChIL-18 by PCR, and was subcloned into Pichia pastoris expression vector pPICZalphaA to construct the recombinant plasmid pPICZalphaA-ChIL-18. After identified by restriction enzymes digestion analysis, PCR and DNA sequencing, the recombinant plasmid was transformed into Pichia pastoris X-33.Then choosing the multi-copy recombinant strains to be induced for expression.Then the bioactivity of rchIL-18 was analysed by Western blot, ELISA and MTT after purified by Sephadex G-100 column.
  
  
  RESULTS
  The chicken IL-18 with the immunogenicity was secreted by Pichia pastoris. It could induce T lymphocytes proliferation and secreting IFN-gamma in vitro.
  
  
  CONCLUSIONS
  The chicken IL-18 with obvious biological activity is secreted by Pichia pastoris X-33.
authors: Animal Science and Technology College, Henan University of Science and Technology, Luoyang 471003, China.
authors: Min
authors: M
authors: Zhao
authors: Chun-Jie
authors: CJ
authors: Zhang
authors: Yin-Ju
authors: YJ
authors: Li
authors: Ting-Cai
authors: TC
authors: Wu
authors: Xiang-Chao
authors: XC
authors: Cheng
authors: Yi-Chen
authors: YC
authors: Liu
authors: Xiao-Li
authors: XL
authors: Yu
authors: Yong-Hua
authors: YH
authors: Qian
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002645:Chickens / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D020382:Interleukin-18 / Q000378:metabolism; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D020382:Interleukin-18; D011994:Recombinant Proteins