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粤黄鸡IL-15基因真核表达及其多克隆抗体的制备
Eukaryotic Expression of Yuehuang Chicken Interleukin 15 Gene and Preparation of Its Polyclonal Antibody
【摘要】 本研究参照GenBank登录的鸡白细胞介素15(ChIL-15)基因序列自行设计一对特异性引物,以RT-PCR、基因克隆,重组真核表达质粒构建、表达及试验兔免疫等技术,成功获得粤黄鸡IL-15基因:全长564bp,编码187个氨基酸,与GenBank上已发表的其它品种鸡IL-15基因序列及其推导的氨基酸序列相似性为99.1%~100%;成功构建鸡IL-15基因重组真核表达质粒pcDNA-ChIL-15;成功制备兔抗鸡IL-15多克隆抗体。为进一步研究粤黄鸡IL-15的生物学功能,试验pcDNA-ChIL-15作鸡的各种新型DNA疫苗的分子免疫佐剂等奠定基础。
【Abstract】 A pair of specific primers was designed according to the published sequences of interleukin 15 (IL-15) of chicken in GenBank. Interleukin 15 gene was amplified by RT-PCR from splenic lymphocytes of Yuehuang chicken , the IL-15 gene of Yuehuang chicken was 564 bp in length , encoding 187 amino acids. The amino acid homologies were 99.1% to 100% compared with the the reported IL-15 of chicken. The gene was then cloned into the expression vector pcDNA3.1 to construct plasmid pcDNA-ChIL-15. And polyclonal antibody was gained after immunizing rabbits with pcDNA-ChIL-15. These findings will be of benefit to further study the role of IL-15 and discuss immune adjuvant effect of pcDNA-ChIL-15 as a novel DNA vaccine immune adjuvant of chicken.
【Key words】 chicken IL-15 gene; eukaryotic expression; polyclonal antibody;
- 【文献出处】 中国家禽 ,China Poultry , 编辑部邮箱 ,2013年05期
- 【分类号】S831
- 【下载频次】62