梅毒螺旋体TpF1蛋白的克隆表达及鉴定  

Amplification, cloning, and expression of the TpF1 protein of Treponema pallidus

作　　者：闫怡[1] 杨宁[2] 董潇阳[1] 刘旭[1] 哈地利亚·哈斯木 周珊[1] 王晓东[1] YAN Yi;YANG Ning;DONG Xiao-yang;LIU Xu;HADILIYA Hasimu;ZHOU Shan;WANG Xiao-dong(Department of Dermatology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830051,China)

机构地区：[1]新疆医科大学第一附属医院皮肤科,新疆乌鲁木齐830054 [2]新疆医科大学第一附属医院临床研究院

出　　处：《中国病原生物学杂志》2021年第10期1134-1138,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81560339)。

摘　　要：目的构建梅毒螺旋体TpF1n基因原核表达载体,表达重组TpF1n,为梅毒螺旋体感染与宿主炎症反应研究提供候选蛋白。方法采用全基因合成法构建原核表达载体pET-28a-TPF1n,表达重组TpF1蛋白,用重组TpF1蛋白刺激人单核巨噬细胞系THP-1,通过ELISA方法检测IL-1β的分泌水平变化。结果成功构建了pET-28a-TPF1n原核表达载体,转染大肠埃希菌后稳定表达约25 ku大小的重组TpF1蛋白。与空白组相比,TpF1蛋白干预组的IL-1β分泌水平显著升高。结论重组TpF1蛋白可显著诱导THP-1细胞分泌IL-1β,提示TpF1蛋白可作为梅毒螺旋体感染引起的宿主炎症反应研究中的候选蛋白,具有重要的应用价值。Objective To express the TpF1 n protein of Treponema pallidum via a prokaryotic expression vector in order to provide a candidate protein for further research on the mechanism of infection and host inflammatory response to T. pallidum. Methods Whole gene synthesis was used to construct the prokaryotic expression vector pET-28 A-TPF1 n. The recombinant TpF1 protein was expressed in the vector. Subsequently, the expressed TpF1 protein was used to stimulate THP-1 cells, which represent a model of human monocyte-derived macrophages. The level of IL-1β secretion was detected using ELISA. Results The prokaryotic expression vector pET-28 A-TPF1 n was successfully constructed. The TpF1 recombinant protein was stably expressed in E. coli and the molecular weight of the TpF1 protein was about 25 ku. Compared to the blank group, the level of IL-1β secretion in the TpF1 intervention group was significantly higher(P<0.01). Conclusion The recombinant TpF1 protein significantly up-regulated the level of IL-1β secretion in THP-1 cells, suggesting that the TpF1 protein warrants use as a candidate protein for the study of the host inflammatory response to a T. pallidum infection.

关 键 词：梅毒螺旋体 炎症 重组抗原 TpF1蛋白 单核巨噬细胞 

分 类 号：R377[医药卫生—病原生物学]