[Expression and analysis of recombinant chicken IL-18 in Pichia pastoris.].
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.
Zhao
Min
M
Animal Science and Technology College, Henan University of Science and Technology, Luoyang 471003, China.
Zhang
Chun-Jie
CJ
Li
Yin-Ju
YJ
Wu
Ting-Cai
TC
Cheng
Xiang-Chao
XC
Liu
Yi-Chen
YC
Yu
Xiao-Li
XL
Qian
Yong-Hua
YH
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
D020382:Interleukin-18; D011994:Recombinant Proteins