猪白细胞介素-6在巴斯德毕氏酵母(Pichia pastoris)中的分泌表达  

Expression of porcine interleukin-6 inPichia pastoris

作　　者：严琳[1] 顾贫[1] 陈焕春[1] 

机构地区：[1]华中农业大学动物医学院,武汉430070

出　　处：《微生物学报》2005年第3期387-391,共5页Acta Microbiologica Sinica

基　　金：国际科学基金-瑞典 (IFS)资助 (B3 5 2 8-1)~~

摘　　要：白细胞介素6 (Interleukin_6 ,IL_6 )是一种具有多种生物学效应的细胞因子,在疾病诊断与疫苗佐剂领域有广阔的应用前景。在本试验中,猪白细胞介素_6 (pIL_6 )的cDNA序列被克隆入甲醇酵母(Pichiapastoris)分泌表达载体pPIC9K中,并转化入P .pastorisGS115菌株。其重组菌株GS115 pPIC9K_IL6经1%甲醇诱导后,能分泌表达分子量约为2 4 5KD的重组蛋白,Westernblot确证为pIL_6。该酵母表达产物无N端糖基化修饰。用依赖IL6生长的B9细胞株检测提纯后的pIL_6 ,其生物学活性可达8×10 4 IU mg。As a kind of pleiotropic cytokine, Interleukin_6 (IL_6) becomes more and more popular in the fields of disease diagnosises and candidate adjuvants. In this study, the porcine Interleukin_6 (pIL_6) cDNAs un_coding singnal peptides was cloned into pPIC9K(the expression vector ofPichia pastoris)to construct the pPIC9K_IL6, which was further transformed into P. pastorisGS115 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. ThisP. pastoris product wasn′t modified by N_hyperglycosylation. After purified, pIL_6 could reach to 8×10 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.

关 键 词：猪白细胞介素6 PICHIA PASTORIS 酵母表达 生物学活性 

分 类 号：Q786[生物学—分子生物学]