| 鹅IL-18基因的克隆和原核表达及其多克隆抗体的制备 |
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| 引用本文： | 郑海华,韩先干,尹会方,周蕾,柯艳坤,黄青云. 鹅IL-18基因的克隆和原核表达及其多克隆抗体的制备[J]. 中国兽医科学, 2010, 40(3) |
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| 作者姓名： | 郑海华 韩先干 尹会方 周蕾 柯艳坤 黄青云 |
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| 作者单位： | 华南农业大学,兽医学院,广东,广州,510642 |
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| 基金项目： | 广东省自然科学基金项目 |
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| 摘 要： | 参考GenBank中鸡、火鸡和鸭IL-18基因序列,设计了1对特异引物,以提取的鹅脾细胞总RNA为模板,应用RT-PCR扩增、克隆鹅IL-18基因中间的333bp序列。序列分析结果表明,其与鸭IL-18基因对应序列相似性最高,达99%。参考鸭IL-18全基因序列设计上、下游引物各3条,以梯度PCR探索扩增、克隆鹅IL-18全基因。以上述策略成功扩增、克隆了3个广东地方杂交品种鹅的IL-18基因。序列分析表明,这3个杂交品种鹅的IL-18基因序列一致,长642bp,含一个603bp的ORF,编码200个氨基酸,分子质量为23.2ku。ORFN端的30个氨基酸为前导序列,其余170个氨基酸为功能蛋白序列。在GenBank中注册了第一条鹅的IL-18基因序列,登录号为EF159728。随后,构建了鹅IL-18基因ORF的重组原核表达质粒pET-GIL-18,经诱导表达、纯化获得重组鹅IL-18融合蛋白,并以其制备了兔抗重组鹅IL-18多克隆抗体。
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| 关 键 词： | 鹅 IL-18基因 克隆 原核表达 多克隆抗体 |
Cloning and prokaryotic expression of goose interleukin-18 gene and the preparation of polyclonal antibody to recombinant goose IL-18 |
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| ZHENG Hai-hua,HAN Xian-gan,YIN Hui-fang,ZHOU Lei,KE Yan-kun,HUANG Qing-yun. Cloning and prokaryotic expression of goose interleukin-18 gene and the preparation of polyclonal antibody to recombinant goose IL-18[J]. Chinese Veterinary Science, 2010, 40(3) |
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| Authors: | ZHENG Hai-hua HAN Xian-gan YIN Hui-fang ZHOU Lei KE Yan-kun HUANG Qing-yun |
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| Affiliation: | ZHENG Hai-hua,HAN Xian-gan,YIN Hui-fang,ZHOU Lei,KE Yan-kun,HUANG Qing-yun(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China) |
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| Abstract: | According to duck,turkey and chicken IL-18 gene sequences available in GenBank,a pair of primers was designed and synthesized to clone the middle sequence of goose IL-18(GIL-18) gene about 333bp by RT-PCR from the total RNA of goose spleen lymphocyte.The sequence analysis showed that the nucleotide homology of the middle sequence between goose and duck was the highest 99%.Then according to the duck IL-18 gene sequence,three upstream and downstream primers were designed and a battery testing of RT-PCR was ca... |
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| Keywords: | goose interleukin-18 cloning prokaryotic expression polyclonal antibody |
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