[Expression and analysis of recombinant pIL-18 in Pichia pastoris].
The porcine IL-18 gene was amplified from recombinant plasmid pGEM-IL-18 by PCR, then the pPIC9K-IL-18 of fusion expression vector was constructed by inserting IL-18 fragment,and was transformed to GS115 by electroporation, multi-copy recombinant strains were screened by G418. The expression of recombinant fusion protein was induced by methanol, SDS-PAGE was used to analyze expression product, fusion protein was purified by Sephadex G-100 column, bioactivity of IL-18 was measured by MTT assays. Experiment results show fusion protein of pIL-18 secreted by GS115,expression reaches the secretion peak of 160 mg/L at 72 h. We have expressed and purified successfully the recombinant pIL-18 with obvious biological activity in Pichia pastoris.
Song
Shi-Bin
SB
Key Laboratory of Veterinary Porasitology of Gansu Province/State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute, Chinese Acadarny of Agricultural Science, Lanzhou 730046, China.
Jing
Zhi-Zhong
ZZ
Chen
Guo-Hu
GH
Wang
Xiao-Xiao
XX
Zong
Rui-Qian
RQ
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D018274:Electroporation; D020382:Interleukin-18 / Q000096:biosynthesis* / Q000235:genetics; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D013552:Swine
D020382:Interleukin-18; D011993:Recombinant Fusion Proteins