猪IL-21的真核表达及其与CD40L共刺激抗体分泌的活性研究  

Eukaryotic expression of porcine interleukin-21 and activity study of IL-21-induced IgG production together with CD40L

作　　者：杨贝贝 符芳[1] 薛美[1] 冯力[1] 刘平黄[1] YANG Bei-bei;FU Fang;XUE Mei;FENG Li;LIU Ping-huang(State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150069,China)

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室

出　　处：《中国预防兽医学报》2019年第5期501-504,533,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家重点研发计划资助(2017YFD0502200)

摘　　要：为研究猪白介素21(IL-21)的生物学功能,本研究在猪IL-21基因(456 bp)的3’端加上猪Ig G Fc标签序列后,将其克隆至真核表达载体pcDNA3.1(-)中,构建重组质粒pcDNA-IL-21-Fc。将该重组质粒转染293T细胞瞬时表达后,利用protein A亲和纯化介质纯化转染上清中的目的蛋白,并检测其纯度与生物学活性。结果显示,构建的重组质粒pcDNA-IL-21-Fc能够在293T细胞中表达相对分子量约为44 ku的猪IL-21融合蛋白;亲和纯化后的猪IL-21融合蛋白能够与CD40L协同呈剂量依懒性地刺激猪外周血淋巴细胞(PBMCs)分泌Ig G。本研究为进一步探究猪IL-21在适应性免疫应答中的作用提供依据。To investigate the biological function of porcine interleukin-21(IL-21),the porcine IL-21 sequence(456 bp)fused with porcine Ig G Fc tag at its 3’end was inserted into pcDNA3.1(-)to construct the recombinant plasmid pcDNA3.1(-)/IL-21-Fc.This recombinant plasmid was subsequently transfected into 293 T cells for transient expression and the recombinant porcine IL-21 fusion protein was purified using protein A affinity chromatography,the purity and biological activity of purified IL-21 fusion protein was determined.Results showed that recombinant plasmid pcDNA3.1(-)/IL-21-Fc was constructed and porcine IL-21 fusion protein with molecular mass of 44 ku was expressed in 293 T cells;The preliminary activity analysis of the purified recombinant porcine IL-21 protein demonstrated that the IL-21 in concert with CD40 L stimulated Ig G secretion in porcine peripheral blood lymphocytes(PBMCs)in a dose-dependent manner,indicating that the recombinant IL-21 has the biological activity.Our study provides a robust tool for further explorations of porcine IL-21 role in adaptive immune responses.

关 键 词：猪IL-21 真核表达 蛋白纯化 抗体分泌 

分 类 号：S852.4[农业科学—基础兽医学]