约氏疟原虫SAP1重组DNA疫苗的构建及鉴定  

Construction and identification of a recombinant DNA vaccine expressing the truncated SAP1 gene of the 17XL strain of Plasmodium yoelii

作　　者：赵佳[1] 左林[2] 罗恩杰[3] 姜小建[1] 

机构地区：[1]西安市中心医院检验科,陕西西安710003 [2]第四军医大学唐都医院放射科,陕西西安710038 [3]中国医科大学病原生物学教研室,辽宁沈阳110013

出　　处：《中国病原生物学杂志》2016年第11期1014-1017,共4页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.30972572)

摘　　要：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 expressing the truncated SAP1 gene of the17 XL strain of Plasmodium yoelii. Methods The truncated SAP1 gene was predicted and analyzed using bioinformatics software and amplified using PCR.The amplified gene was cloned into the eukaryotic expression vector pcDNA3.1（＋）,and the recombinant expression vector pcDNA3.1（＋）/SAP1 was constructed.COS-7cells were transfected with the recombinant expression vector and the expression of the recombinant protein was observed with indirect immunofluorescence.The recombinant protein was detected with SDS-PAGE and Western blotting. Results The truncated SAP1 gene was successfully amplified using PCR.The eukaryotic expression vector pcDNA3.1（＋）/SAP1 was constructed and expressed transiently in vitro.The product of expression reacted specifically with polyclonal antiserum. Conclusion The recombinant DNA vaccine constructed here was expressed transiently in mammalian cells and the expressed protein was immunoreactive.The immunoprotective action of the vaccine needs to be studied further.

关 键 词：疟原虫 约氏 SAP1 DNA疫苗 基因表达 

分 类 号：R382.31[医药卫生—医学寄生虫学]