牛IFN-ε基因克隆、表达与抗病毒活性分析  

Cloning, expression and antiviral activity analysis of bovine IFN-ε

作　　者：王庭柱[1,2] 郭永丽[1] 高明春[1] 罗修鑫 安东[1] 刘莹[1] 师东方[1] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]黑龙江省农垦总局建三江管理局八五九农场,黑龙江双鸭山156326

出　　处：《东北农业大学学报》2015年第12期39-44,共6页Journal of Northeast Agricultural University

基　　金：现代农业(奶牛)产业技术体系项目(CARS-37);国家科技支撑计划项目(2012BAD12B05;2012BAD12B03)

摘　　要：为分析预测牛IFN-ε基因功能及特性,研究首先从牛肝基因组中克隆该基因,推导牛IFN-ε基因对应氨基酸并作生物信息学分析,在m RNA水平验证牛IFN-ε基因为可转录基因。构建包含牛IFN-ε成熟肽基因的原核表达载体p ET32a-Bo NE,将重组载体转移至大肠杆菌Rosetta TM(DE3)p Lys S,IPTG诱导表达,融合蛋白r Bo NE主要以包涵体形式存在。结果表明,经MDBK-VSV干扰素活性系统检测,牛IFN-ε融合蛋白具有抗病毒活性。组织分布表明,其可在肝、肾、胸腺、小肠以及睾丸中组成性表达,但心脏未见表达。In order to analyze the functions and characterization of predicted bovine IFN-ε gene, a novel bovine IFN-ε gene was cloned from bovine liver genome, the corresponding amino acids were deduced and the characterization was analyzed with bioinformatics software, and bovine IFN-ε was verified to be the transcribed gene at the mRNA level. Then the prokaryotic expression vector pET32a-BoNE containing the mature peptide gene of bovine IFN-ε was constructed, and the recombinant vector was transformed into E. coil RosettaTM （DE3） pLysS. After IPTG induction, the fusion protein rBoNE mainly expressed as the insoluble form. The results showed antiviral activity was analyzed by MDBK-VSV system, the result revealed that bovine IFN-ε had antiviral activity. Tissue distribution showed that bovine IFN-ε could be expressed constitutively in liver, kidney, thymus, small intestine and testis, but not expressed in the heart.

关 键 词：牛IFN-ε 克隆表达 融合蛋白 抗病毒活性 

分 类 号：S852.42[农业科学—基础兽医学]