| 鸡IL-18重组真核表达载体的构建及其对ND灭活苗的佐剂作用 |
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| 引用本文： | 陈红英,李新生,金钺,张红英,文英会,崔保安.鸡IL-18重组真核表达载体的构建及其对ND灭活苗的佐剂作用[J].江西农业大学学报,2009,31(5). |
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| 作者姓名： | 陈红英 李新生 金钺 张红英 文英会 崔保安 |
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| 作者单位： | 1. 河南农业大学,牧医工程学院,河南,郑州,450002;河南省动物性食品安全重点实验室,河南,郑州,450002
2. 河南农业大学,牧医工程学院,河南,郑州,450002 |
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| 基金项目： | 国家"十一五"科技支撑计划专项 |
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| 摘 要： | 从pGEM-ChIL-18克隆质粒扩增获得鸡IL-18(ChIL-18)全基因片段,并将其重组到真核表达载体pcDNA3.1(+)中,构建重组真核表达质粒pcDNA3.1/ChIL-18(简称pChIL-18).经PCR、酶切和基因测序证实,所获重组质粒含有目的片段,且连接、构建正确.pChIL-18转染Cos7细胞,转染细胞中含ChIL-18基因的mRNA,表达产物对鸡淋巴细胞具有明显诱导转化作用.pChIL-18对ND灭活苗免疫增强作用的研究表明pChIL-18能够提高ND灭活苗诱发的细胞免疫应答,为后续基因疫苗的研究奠定基础.
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| 关 键 词： | 白细胞介素18 鸡 真核表达 淋巴细胞转化试验 |
Construction of Eukaryotic Expression Plasmids Encoding Chicken IL-18 Gene and Its Immunoenhancement on Newcastle Disease Inactivated Vaccine |
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| CHEN Hong-ying,LI Xin-sheng,JIN Yue,ZHANG Hong-ying,WEN Ying-hui,CUI Bao-an.Construction of Eukaryotic Expression Plasmids Encoding Chicken IL-18 Gene and Its Immunoenhancement on Newcastle Disease Inactivated Vaccine[J].Acta Agriculturae Universitis Jiangxiensis,2009,31(5). |
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| Authors: | CHEN Hong-ying LI Xin-sheng JIN Yue ZHANG Hong-ying WEN Ying-hui CUI Bao-an |
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| Institution: | CHEN Hong-ying1,2,LI Xin-sheng1,JIN Yue1,ZHANG Hong-ying1,WEN Ying-hui1,CUI Bao-an1,2* (1.College of Animal Husbandry and Veterinary,He'nan Agricultural University,Zhengzhou 450002,China,2.Animal Food Safety Key Laboratory,He'nan Province,China) |
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| Abstract: | The whole sequence of chicken IL - 18 ( ChIL - 18) gene was amplified by PCR from plasmid pGEM-ChIL-18 and inserted into eukaryotic expression plasmid pcDNA3. 1( + )to construct recombinant eukaryotic expression plasmid pcDNA3. 1/ChIL - 18 (pChIL - 18). The results of identification of PCR and enzyme digestion and sequencing for recombinant plasmid showed the sequence of the eukaryotic expression plasmid pChIL - 18 in the reading frame and the ligation part were correct. After the transfection of pChIL - 18 into Cos7 ceils,ChIL -18 mRNA expression was found in Cos7 cells. The results of MTT assay showed that the expression of ChIL - 18 protein in Cos7 cell could induce significandy transformation of chicken T lymphocytes. Immunoenhancement effect of the pChIL-18 plasmid on Newcasde disease vaccine was observed by proliferation response of T lymphocytes from spleen. It can obviously enhance the cell - mediated immune response. |
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| Keywords: | interleukin -18 chicken eukaryotic expression lymphocyte transformation test |
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