摘要
为对犬孢子虫MAG1与IFN-γ融合基因重组真核质粒进行免疫学评价,本实验以含有MAG1-IFN-γ基因片段的克隆质粒为模板,扩增MAG1-IFN-γ目的片段,构建pVAX-MAG1-IFN-γ重组真核表达质粒,将其转染Vero细胞,采用间接免疫荧光(IFA)和western blot技术检测MAG1基因在Vero细胞中的表达,并免疫BALB/c小鼠,进行免疫学评价。结果表明,PCR扩增获得基因片段大小为1 536 bp,与GenBank中登录的MAG1和IFN-γ核苷酸序列的同源性为99%;IFA检测MAG1基因在Vero细胞中获得瞬时表达,转染的Vero细胞呈现特异性绿色荧光;western blot分析表达蛋白的分子量为57 ku,具有较好的反应原性。将构建的重组真核表达质粒免疫BALB/c小鼠,经间接ELISA和T淋巴细胞亚群含量测定表明,重组质粒具有较好的免疫原性。本实验为新孢子虫病核酸疫苗的深入研究奠定了基础。
To evaluate immune responses of the recombinant plasmid co-expressing MAGI gene of Neospora caninum and IFN-y, the recombinant plasmid of pVAX-MAGI-IFN-y was constructed and transfected into Vero cells. The expression of MAGI gene was identified by indirect immunofluorescent assay and western blot. The results showed that MAGI-IFN-y gene was transiently expressed in Vero cells and the protein expressed in Vero cells had good reactogenicity. Furthermore, BALB/c mice were immunized by pVAX-MAGI-IFN-y Indirect ELISA and determination of T lymphocyte subsets showed that the recombinant plasmid was able to induce efficient immune responses in inoculated mice. These results laid the foundation for the further research of DNA vaccine against Neospora caninum.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2012年第8期662-666,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
国家自然科学基金(31160501)
