具抗病毒活性的重组牛IFN-αA的冷激诱导表达与鉴定  

Expression and identification of recombinant BoIFN-αA with the antiviral activity using intein-cold shock induced expression system

作　　者：孙旭燕[1] 崔子寅[1] 高明春[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2013年第3期227-231,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技支撑计划项目(2012BAD12B05);国家科技支撑计划子课题(2012BAD12B03-3);现代农业(奶牛)产业技术体系(CARS-37);黑龙江省科技攻关项目(GA09B302)

摘　　要：为了在E.coli表达系统中高效表达具有抗病毒活性的重组牛α干扰素蛋白(rBoIFN-α),本研究通过PCR扩增牛α干扰素A亚型(BoIFN-αA)基因,并在其上游连接内含肽(SDI)序列,克隆于pColdⅢ中构建内含肽-冷激表达重组质粒pColdⅢ-NusA-SDI-BoIFN-α,16℃低温诱导表达。结果表明,rBoIFN-α实现了高效表达,表达产物主要以可溶形式存在,经Ni柱纯化后得到浓度为0.12 mg/mL的目的蛋白。经MDBK-BVDV干扰素活性检测系统检测显示,表达的rBoIFN-α抗病毒活性为9.86×105U/mg。本研究结果为具有抗病毒活性的rBoIFN-α后续应用研究奠定了基础。To improve the expression of soluble recombinant bovine interferon-α-A （BolFN-αA） in Escherichia coli, BolFN-αA gene was fused with the Ssp DnaB mini-intein （SDI） at the upstream of BolFN-α cDNA and cloned into the expression vector pColdⅢ to construct pColdⅢ-NusA-SDI-BolFN-α, which was transformed into E.coli and induced by IPTG at 16 ℃. The result showed that the fusion protein was expressed in soluble form. Furthermore, the purified protein showed the antiviral activity in MDBK-BVDV system and the antiviral activities were 9.86 ×10^5 U/mg. In conclusion, the expression of BolFN-α lay a foundation for the further study and application.

关 键 词：牛IFN-α 冷激 内含肽 可溶性表达 抗病毒活性 

分 类 号：S852.4[农业科学—基础兽医学]