摘要
为构建表达新孢子虫重组蛋白SRS2原核表达系统,本研究以新孢子虫基因组为模板PCR扩增SRS2蛋白部分基因,获得大小约为998bp的目的片段,并克隆到大肠杆菌PET-32a载体中,IPTG诱导重组大肠杆菌,通过SDS-PAGE和免疫印迹分析表达产物并鉴定其生物学活性。经SDS-PAGE分析,表达的重组SRS2蛋白分子量约为54kD,免疫印迹结果表明,表达的重组蛋白与牛抗新孢子虫阳性血清反应形成一条特异性蛋白条带。本实验为进一步研究诊断和治疗新孢子虫病奠定了基础。
To construct recombinant plasmid expressing Neospora caninum SRS2, the SRS2 gene from a Neospora caninum strain was amplified by PCR with a pair of primers designed according to the sequence in GenBank.A 998 bp gene was amplified and cloned into the vector PET-32a.The expression of SRS2 protein was induced by IPTG .SDS-PAGE analysis indicated that the expressed recombinant protein of SRS2 was about 54 kD.The Western blot analysis confirmed the recombinant protein was recognized by positive serum anti- Neospora caninum.The study provided a foundation for further study on the diagnosis and vaccine of neosporosis.
出处
《中国动物检疫》
CAS
2012年第9期43-45,59,共4页
China Animal Health Inspection
