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| 猪IL-15 cDNA真核表达载体的构建、表达及其免疫佐剂效应 | |
| 引用本文： | 何谦,陈钜豪,房红莹,卢俊鹏,张欣,崔敏,曾小娜,刘健,罗满林.猪IL-15 cDNA真核表达载体的构建、表达及其免疫佐剂效应[J].中国兽医学报,2011,31(8). |
| 作者姓名： | 何谦 陈钜豪 房红莹 卢俊鹏 张欣 崔敏 曾小娜 刘健 罗满林 |
| 作者单位： | 1. 暨南大学生命与健康工程研究院,广东广州,510632 2. 广州市工商行政管理局白云分局,广东广州510402／华南农业大学兽医学院动物传染病实验室,广东广州510642 3. 华南农业大学兽医学院动物传染病实验室,广东广州,510642 |
| 摘 要： | 参考GenBank发表猪IL-15mRNA序列设计引物,用RT-PCR方法扩增猪IL-15cDNA,并克隆到pMD18-T载体中,通过PCR、酶切和测序验证克隆正确,再亚克隆到真核表达载体pcDNA-3.1（＋）上,得到重组质粒pcD-NA-pIL-15。在脂质体介导下,重组质粒pcDNA-pIL-15转染AD-293细胞。以鼠抗猪IL-15为一抗,用间接免疫荧光分析表明猪IL-15基因均在AD-293细胞中成功进行了瞬时表达。小鼠免疫试验表明,pcDNA-pIL-15作为免疫佐剂,能够有效提高小鼠的脾T细胞增殖,加强pcDNA-ORF2（PCV2）质粒免疫过程中的特异性中和抗体的产生,为进一步研制猪IL-15基因佐剂疫苗及进行临床实验奠定基础。 |
| 关 键 词： | 猪IL-15 真核表达 瞬时表达 佐剂疫苗 中和抗体水平 |
Construction and expression of porcine interleukin-15 cDNA eukaryotic expression vector and its enhancing of immunity as immunoadjuvants |
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| HE Qian,CHEN Ju-hao,FANG Hong-ying,LU jun-peng,ZHANG Xin,CUI Min,ZENG Xiao-na,LIU Jian,LUO Man-lin.Construction and expression of porcine interleukin-15 cDNA eukaryotic expression vector and its enhancing of immunity as immunoadjuvants[J].Chinese Journal of Veterinary Science,2011,31(8). | |
| Authors: | HE Qian CHEN Ju-hao FANG Hong-ying LU jun-peng ZHANG Xin CUI Min ZENG Xiao-na LIU Jian LUO Man-lin |
| Institution: | HE Qian1,CHEN Ju-hao2,3,FANG Hong-ying3,LU jun-peng3,ZHANG Xin3,CUI Min3,ZENG Xiao-na3,LIU Jian3,LUO Man-lin3(1.Institute of Life and Health Engineering,JiNan University,Guangzhou 510632,China,2.Baiyun Branch,Administration for Industry and Commerce of Guangzhou,Guangzhou 510403,3.College of Veterinary Medicine,South China Agricultural University,Guangzhou 510640,China) |
| Abstract: | According to the sequences of porcine interleukin-15 messenger ribonucleic acid(pIL-15mRNA) published in GenBank,the primers were designed to amplify IL-15 cDNA,which was cloned to into the pMD18-T vector.The product was proved correctly by PCR,enzyme digestion and sequencing,and subcloned into the eukaryotic expression vector pcDNA-3.1(+),then the recombinant plasmid pcDNA-pIL-15 was transfected to AD-293 cells by the liposome-mediated.With mouse anti-pig IL-15 as the first antibody,indirect immunofluoresc... |
| Keywords: | porcine Interleukin-15 eukaryotic expression transient expression adjuvant vaccine |
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