| 鸡IL-18成熟蛋白基因原核表达载体的构建及多克隆抗体的制备 |
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| 引用本文： | 刘一尘,张春杰,程相朝,张谦,李银聚,吴庭才,赵战勤.鸡IL-18成熟蛋白基因原核表达载体的构建及多克隆抗体的制备[J].河南农业科学,2009(2). |
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| 作者姓名： | 刘一尘 张春杰 程相朝 张谦 李银聚 吴庭才 赵战勤 |
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| 作者单位： | 1. 河南科技大学,动物科技学院,河南,洛阳,471003
2. 郑州后羿集团,河南,郑州,450002 |
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| 摘 要： | 应用RT-PCR技术得到鸡IL-18成熟蛋白基因(chicken mature interleukin-18),将克隆片段与原核表达载体pET28a+连接,构建了重组表达载体pET28a+-ChMIL18,转化至大肠杆菌BL21感受态细胞,获得重组表达菌株。经IPTG诱导表达,SDS-PAGE分析后,用含有融合蛋白的聚丙烯酰胺凝胶电泳颗粒免疫家兔,制备兔抗鸡IL-18多克隆抗体,Western-Blot检测抗体效价和特异性。结果表明,鸡IL-18在BL21中得到了表达,相对分子质量约为23kD,获得了具有较高效价的特异性多克隆抗体。为进一步研究鸡IL-18成熟蛋白的生物学活性和作用机制奠定了基础。
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| 关 键 词： | 鸡IL-18成熟蛋白基因 原核表达 多克隆抗体 |
Prokaryotic Expression of Chicken Mature Interleukin 18 Gene and Preparation of Polyclonal Antibodies |
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| LIU Yi-chen,ZHANG Chun-jie,CHENG Xiang-chao,ZHANG Qian,LI Yin-ju,WU Ting-cai,ZHAO Zhan-qin.Prokaryotic Expression of Chicken Mature Interleukin 18 Gene and Preparation of Polyclonal Antibodies[J].Journal of Henan Agricultural Sciences,2009(2). |
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| Authors: | LIU Yi-chen ZHANG Chun-jie CHENG Xiang-chao ZHANG Qian LI Yin-ju WU Ting-cai ZHAO Zhan-qin |
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| Institution: | 1.College of Animal Science and Technology;Henan University of Science and Technology;Luoyang 471003;China;2.Zhengzhou Hou Yi Group;Zhengzhou 450002;China |
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| Abstract: | The gene of chicken mature interleukin-18(ChMIL-18) was amplified by RT-PCR.The prokaryotic expression vector pET28a+-ChMIL 18 was constructed,transformed into E.coli BL21 and induced by IPTG.The fusion proteins were isolated and used for the preparation of the polyclonal antisera against ChMIL-18 by immunizing the New Zealand rabbits.Western-Blot was performed using the purified polyclonal antibodies the results showed that the protein ChMIL-18 was successfully expressed in BL21 and the molecular weigh was... |
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| Keywords: | Chicken mature interleukin-18 gene Prokaryon expression Polyclonal antibody |
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