[Expression of porcine interleukin-6 in Pichia pastoris].
As a kind of pleiotropic cytokine, Interleukin-6 (IL-6) becomes more and more popular in the fields of disease diagnosis and candidate adjuvants. In this study, the porcine Interleukin-6 (pIL-6) cDNAs un-coding signal peptides was cloned into pPIC9K (the expression vector of Pichia pastoris) to construct the pPIC9K-IL6, which was further transformed into P. pastoris GS115 strain. After being induced by 1% methanol, the recombinant strain excreted a protein in supernate of culture, with molecular weight of 24.5KD approximately, which was proved to be pIL6 through SDS-PAGE and western blotting. This P. pastoris product wasn't modified by N-hyperglycosylation. After purified, pIL-6 could reach to 8 x 10(4) IU/mg of bioactivity unit by the cell proliferation of B9 assay. Through researching the characters of the recombinant protein pIL6, we have made a great progress on the study of rpIL6 as veterinary vaccine's adjuvant.
Yan
Lin
L
Laboratory of Animal Virology, College of Veterinary Medicine, Huazhong Agriculture University, Wuhan 430070, China. yanlinalin@yahoo.com.cn
Gu
Pin
P
Chen
Huan-chun
HC
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000235:genetics; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013552:Swine
D018076:DNA, Complementary; D015850:Interleukin-6; D011994:Recombinant Proteins