犬干扰素α1和胸腺肽α1融合蛋白在毕赤酵母GS115中的表达  

Expression of canine IFNα1-THYα1 fusion protein in pichia pastoris GS115

作　　者：田春玲 马玉忠[1] 秦建华[1] 汪恩强[1] TIAN Chun-ling;MA yu-zhong;QIN Jian-hua;WANG En-qiang(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China)

机构地区：[1]河北农业大学动物医学院,河北保定071001

出　　处：《河北农业大学学报》2018年第1期88-91,105,共5页Journal of Hebei Agricultural University

基　　金：河北省现代化农业产业技术体系奶牛产业创新团队建设项目(1004023)

摘　　要：为探索犬干扰素和胸腺肽融合蛋白的抗病毒活性,根据已知的基因序列,合成了犬干扰素(IFNα1)和胸腺肽(THYα1)的基因片段,通过一个连接肽将IFNα1和THYα1片段连接起来。将该基因克隆于pPIC9K载体中,构建重组质粒pPIC9K-IFNα1-THYα1,转化到感受态细胞毕赤酵母GS115受体菌中,30℃培养18h至菌体OD600nm值约为4时用甲醇诱导培养5d,使最终浓度维持1%,经SDS-PAGE和Western-blot鉴定证实,获得高纯度的重组蛋白。以MDCK/VSV抗病毒检测系统对目的蛋白进行体外活性检测,结果显示该蛋白具有抗病毒活性。In order to investigate the antiviral activity of the fusion protein of Canine interferon and thymosin,the gene fragments of Canine interferon-α1(IFNα1)and thymosinα1(THYα1)were synthesized and ligated by a linked peptide based on the known gene sequences of Canine interferon-α1(IFNα1)and thymosinα1(THYα1).The gene was cloned into pPIC9K vector,and the recombinant plasmid pPIC9K-IFNα1-THYα1 was constructed.The recombinant plasmid was transferred into Pichia Pastoris receptive cell GS115 and cultured at 30℃for 18 hours.The cultured plasmid was induced by methanol for 5 days at the OD 600nm value of 4 to make the final concentration to 1%.Based on SDS-PAGE and Western-blot results,the recombinant protein of Canine IFNα1-THYα1 with high-purity was obtained successfully.In vitro activity detection of target protein by MDCK/VSV virus detection system indicates that the protein has antiviral activity.

关 键 词：犬干扰素α1 胸腺肽Α1 融合蛋白 

分 类 号：S855.3[农业科学—临床兽医学]