中国兽医科学2017，Vol.47Issue(3)：328-333,6.DOI:10.16656/j.issn.1673-4696.2017.03.009
莫氏巴贝斯虫trap基因的克隆表达及反应原性分析
Cloning, expression and immunoreactivity analysis of Babesia motasi trap gene
摘要
Abstract
In the present study,focused on the Babesia motasi Lintan,the full length of trap gene was amplified from genomic DNA and cDNA.The structures of the gene/protein were analyzed and verified by using bioinformatic softwares.A prokaryotic expression vector,pET-30a-trap,was constructed and transformed into Escherichia coli BL21(DE3) pLysS to induce protein expression.Supernatant and precipitate were analyzed by SDS-PAGE after the expression product was lysed by ultrasonication.Soluble rBmTRAP was purified and then its immunoreactivity was determined by western-blot.The result showed that the trap gene contained 4 introns and 5 exons with an open reading frame(ORF) of 2100 bp.The 1-23 animo acids were a signal peptide sequence,45-201 and 238-302 were vWAand TSP1 motifs of TRAP protein family,respectively.The rBmTRAPwas expressed as two forms,soluble protein and insoluble inclusion bodies.The rBmTRAPcould be specifically recognized by positive sera from sheep infected by B.motasi Lintan/Tianzhu.No cross-reactions was present with the sera of B.motasi Hebei/Ningxian,Babesia sp.Xingjiang/Dunhuang,Theileria luwenshuni and Anaplasma ovis.The result suggests that rBmTRAP has the potetial for considering as a candidate of antigen used in sero-diagnostic methods.It was established a foundation for developing immulogical diagnostic methods for differentiating infection of B.motasi Lintan/Tianzhu from other ovine parasites in future.关键词
莫氏巴贝斯虫/trap基因/原核表达/反应原性分析Key words
Babesia motasi/trap/prokaryotic expression/immunoreactivity analysis分类
农业科技引用本文复制引用
何欣,刘军龙,刘爱红,王锦明,牛庆丽,李有全,殷宏,罗建勋,关贵全..莫氏巴贝斯虫trap基因的克隆表达及反应原性分析[J].中国兽医科学,2017,47(3):328-333,6.基金项目
国家重点基础研究发展计划(973)项目(2015CB150300) （973）
国家自然科学基金项目(31072130) （31072130）
农业科技创新工程项目(ASTIP) （ASTIP）
甘肃省青年科技基金项目(145RJYA272) （145RJYA272）
