| 猪干扰素α-1基因的原核表达与多克隆抗体制备 |
| |
| 引用本文： | 刘娜,李春秋,高晶,耿雨菲,王欣宇,苏明俊,魏姗,王志慧,王建发,武瑞,孙东波.猪干扰素α-1基因的原核表达与多克隆抗体制备[J].畜牧与饲料科学,2014(12):1-4. |
| |
| 作者姓名： | 刘娜 李春秋 高晶 耿雨菲 王欣宇 苏明俊 魏姗 王志慧 王建发 武瑞 孙东波 |
| |
| 作者单位： | 黑龙江八一农垦大学动物科技学院,黑龙江 大庆,163319 |
| |
| 基金项目： | 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。
|
| 关 键 词： | 猪干扰素α-1 原核表达 抗体制备 |
Prokaryotic Expression of Porcine Interferonα-1 and Preparation of Polyclonal Antibodies |
| |
| LIU Na,LI Chun-qiu,GAO Jing,GENG Yu-fei,WANG Xin-yu,SU Ming-jun,WEI Shan,WANG Zhi-hui,WANG Jian-fa,WU Rui,SUN Dong-bo.Prokaryotic Expression of Porcine Interferonα-1 and Preparation of Polyclonal Antibodies[J].Animal Husbandry and Feed Science,2014(12):1-4. |
| |
| Authors: | LIU Na LI Chun-qiu GAO Jing GENG Yu-fei WANG Xin-yu SU Ming-jun WEI Shan WANG Zhi-hui WANG Jian-fa WU Rui SUN Dong-bo |
| |
| Institution: | (College of Animal Science and Technology, Heilongjiang Bayi Agricultural University, Daqing 163319, China) |
| |
| Abstract: | 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. |
| |
| Keywords: | porcine interferonα-1 prokaryotic expression preparation of antibodies |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
