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目的构建含有约氏疟原虫SAP1(sporozoite asparagine-rich protein 1)截短基因的重组DNA疫苗,并进行鉴定。方法应用生物学信息软件预测分析并扩增SAP1截短基因,将该基因克隆入真核表达载体pcDNA3.1(+)中,构建重组表达载体pcDNA3.1(+)/SAP1。将重组载体转染COS-7细胞,进行体外瞬时表达并进行SDS-PAGE和Western blot鉴定。结果成功扩增SAP1截短基因并构建含有该基因的真核表达载体pcDNA3.1(+)/SAP1,其体外瞬时表达产物能与多克隆抗血清发生特异性结合反应。结论构建的重组DNA疫苗可在哺乳动物细胞中瞬时表达,表达的蛋白具有免疫反应性,其免疫保护作用有待进一步研究。
Objective To construct and identify a recombinant DNA vaccine containing the truncated gene of SAP1 (sporozoite asparagine-rich protein 1). METHODS: The truncated gene of SAP1 was predicted and amplified by using biological information software. The gene was cloned into eukaryotic expression vector pcDNA3.1 (+) to construct recombinant expression vector pcDNA3.1 (+) / SAP1. The recombinant vector was transfected into COS-7 cells for transient expression in vitro and identified by SDS-PAGE and Western blot. Results The truncated gene of SAP1 was successfully amplified and the eukaryotic expression vector pcDNA3.1 (+) / SAP1 containing this gene was constructed. The transient expression product of the recombinant plasmid was able to specifically react with polyclonal antiserum. Conclusion The constructed recombinant DNA vaccine can be transiently expressed in mammalian cells. The expressed protein is immunoreactive and its immunoprotection remains to be further studied.