牛源犬新孢子虫MAG1基因的克隆及在Vero细胞中表达

摘要

为了解牛源犬新孢子虫MAG1基因的生物学特性,本实验应用PCR技术扩增牛源犬新孢子虫MAG1基因,构建真核表达重组质粒pVAX-MAG1,将鉴定正确的pVAX-MAG1重组质粒转染Vero细胞,应用间接荧光检测方法(1FA)和western blot技术检测MAG1基因在Vero细胞中的表达.结果显示,扩增的牛源犬新孢子虫MAG1基因长度为1047bp,与GenBank中登录的MAG1 (EF580924.1)核苷酸序列同源性为99％,IFA检测MAG1基因在Vero细胞中获得瞬时表达,western blot分析表达蛋白的分子量为39 ku,具有较好的反应原性.本实验为新孢子虫病核酸疫苗与诊断试剂盒的研究奠定了基础.%To inestigate the biological characteristics of bovine Neospora caninum MAGI gene, the gene was amplified by PCR and inserted into eukaryotic expression vector. The resultant recombinant plasmid of pVAX-MAG 1 was transfected into Vero cells and the expression of MAGI was detected by indirect immunofluorescent assay, and western blot showed that the molecular weight of the protein was 39 ku and the protein had a positve reaction with anti MAGI serum. The results could be useful to DNA vaccine development.