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

摘要

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.%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.