猪干扰素α-1基因的原核表达与多克隆抗体制备  

Prokaryotic Expression of Porcine Interferon α-1 and Preparation of Polyclonal Antibodies

作　　者：刘娜[1] 李春秋[1] 高晶[1] 耿雨菲[1] 王欣宇[1] 苏明俊 魏姗[1] 王志慧[1] 王建发[1] 武瑞[1] 孙东波[1] 

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《畜牧与饲料科学》2014年第12期1-4,共4页Animal Husbandry and Feed Science

基　　金：黑龙江省高校新世纪优秀人才项目(1252-NCET-016)

摘　　要：[目的]利用大肠杆菌原核表达系统对猪干扰素α-1(Po IFNα-1)基因进行表达,制备Po IFNα-1重组蛋白多克隆抗体。[方法]根据Gen Bank发表的Po IFNα-1核苷酸序列,合成密码子优化的Po IFNα-1基因,构建Po IFNα-1基因的p ET-28a原核表达载体,转化大肠杆菌BL21(DE3)宿主菌后,利用IPTG进行诱导表达。表达的Po IFNα-1重组蛋白纯化后,免疫昆明系小鼠,制备多克隆抗体。采用Western blot技术检测Po IFNα-1重组蛋白的多克隆抗体与天然Po IFNα-1的反应性。[结果]合成的Po IFNα-1基因在大肠杆菌BL21(DE3)宿主菌中成功表达,针对Po IFNα-1重组蛋白的多克隆抗体能够识别天然的猪干扰素α-1。[结论]合成的Po IFNα-1基因在大肠杆菌中获得了成功表达,为后续研究奠定了基础。Objective] The porcine interferon alpha 1 gene （PoIFNα-1） was expressed by using Escherichia coli prokaryotic expression system, and polyclonal antibodies against the recombinant protein PoIFNα-1 were prepared. [Method] The codon-optimized PoIFNα-1 gene was synthesized according to the nucleotide sequence published by GenBank, and then the synthesized PoIFNα-1 gene was cloned into the vector pET-28a. After transformation of the recombinant plasmid into E. coli BL21 （DE3） host bacteria, the recombinant bacterium was induced by using IPTG. After purification of the recombinant protein PoIFNα-1 expressed in E. coli BL21 （DE3）, the polyclonal antibodies against the recombinant protein PoIFNα-1 was prepared in Kunming mice, and then reactivity of the prepared polyclonal antibodies with native porcine interferon alpha 1 was determined by Western blot. [Result] The synthesized PoIFNα-1 gene was successfully expressed in E. coli BL21 （DE3）, and the polyclonal antibody against recombinant protein PoIFNα-1 could recognize the native porcine interferon-α-1. [Conclusion] The synthesized PoIFNα-1 gene was successfully expressed in E. coli, which provided some basis for following further research of antivirus.

关 键 词：猪干扰素α-1 原核表达 抗体制备 

分 类 号：Q78[生物学—分子生物学] S852.4[农业科学—基础兽医学]