中国兽医科学2017，Vol.47Issue(12)：1503-1509,7.DOI:10.16656/j.issn.1673-4696.2017.12.006
吕氏泰勒虫TlSP无跨膜区重组蛋白的表达及反应原性分析
Expression and reactionogenicity analysis of the recombinant transmembrane region-absent TlSP protein of Theileria luwenshuni
摘要
Abstract
The total RNA of Theileria luwenshuni merozoites was extracted and the TlSP gene of Theileria luwenshuni was successfully amplified by RT-PCR.The epitopes and transmembrane regions of the ~ luwenshuni surface protein (TlSP) were predicted using bioinformatics methods.The TlSP fragment without 1,2 and 3 transmembrane regions was cloned into prokaryotic expression vector pET-30a,respectively,and the recombinant plasmids was transformed into Escherichia coli BL21(DE3)pLysS.The recombinant TlSP was expressed and purified,and its reactionogenicity was analyzed.The results showed that N-terminal of the TlSP had good antigenicity,and three transmembrane regions (185-207 aa,228-250 aa,255-277 aa) were presented at the C-terminal end.The SDS-PAGE result showed that only TlSP without 3 transmembrane regions could be expressed,and the Western-blot showed that the recombinant TlSP had strong reactions with positive serafrom T.luwenshuni,T.uilenbergi,T.ovisand T.annulata,but no cross-reactions with positive sera against other Babesia and Anaplasma ovis,respectively.The study laid the foundation for the study of TlSP candidate subunit vaccines.关键词
吕氏泰勒虫/TlSP基因/跨膜区/原核表达/反应原性分析Key words
Theileria luwenshuni/TlSP gene/transmembrane region/prokaryotic expression/reactionogenicity analysis分类
农业科技引用本文复制引用
韩元,李禤,殷宏,罗建勋,蔡葵蒸,许强,杨学财,李有全,郭鹏飞,关贵全,刘军龙,刘爱红,刘光远..吕氏泰勒虫TlSP无跨膜区重组蛋白的表达及反应原性分析[J].中国兽医科学,2017,47(12):1503-1509,7.基金项目
国家重点研发计划项目(2017YFD0501200,2016YFC1202000,2016YFC1202002) （2017YFD0501200,2016YFC1202000,2016YFC1202002）
肉牛牦牛体系(CARS-37) （CARS-37）
国家自然科学基金项目(31272556) （31272556）
