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| 抗炎、抗凝双效融合蛋白TAP-SSL5表达载体的构建及其功能研究 | |
| 引用本文： | 曲小龙,胡厚源,李敏,黄德兴,梁华.抗炎、抗凝双效融合蛋白TAP-SSL5表达载体的构建及其功能研究[J].第三军医大学学报,2010,32(1). |
| 作者姓名： | 曲小龙 胡厚源 李敏 黄德兴 梁华 |
| 作者单位： | 1. 第三军医大学西南医院心血管内科,重庆市介入心脏病学研究所,重庆,400038 2. 第三军医大学西南医院心血管内科,护理系心理学教研室,重庆,400038 3. 8008 Australia Melbourne,BakerIDI Heart & Diabetes Institute |
| 基金项目： | 国家高技术研究发展计划(863计划,2009AA02Z115);;国家自然科学基金(30770851,30871059)~~ |
| 摘 要： | 目的构建一种具有抗炎和抗凝双效功能的融合蛋白,并对其功能进行初步鉴定。方法将重组金黄色葡萄球菌超抗原样蛋白-5(staphylococcal superantigen-like protein-5,SSL5)与蜱抗凝血肽(tick anticoagulant peptide,TAP)融合,从金黄色葡萄球NCTC8325菌株中克隆SSL5基因;经DNA重组技术,将编码pelB前导序列、SSL5和TAP的基因重组并克隆于原核表达载体pHOG21中,再转染于大肠杆菌BL21中扩增表达,制备融合蛋白SSL5-TAP或TAP-SSL5,采用流式细胞仪检测融合蛋白是否和SSL5一样,具有与粒细胞表面P-选择素糖蛋白配体-1(P-selectin glycoprotein ligand-1,PSGL-1)结合的特性;采用反应底物法在体外检测融合蛋白对活化的凝血因子X(factor Xa,FXa)活性的抑制作用。结果TAP-SSL5保留了SSL5与PSGL-1结合的特性,并且融合蛋白TAP-SSL5可显著抑制FXa的活性(P<0.01),抑制率达39.5%。结论融合蛋白TAP-SSL5是一种以PSGL-1为靶向的新型抗炎、抗... |
| 关 键 词： | 金黄色葡萄球菌超抗原样蛋白-5 蜱抗凝血肽 融合蛋白 P-选择素糖蛋白配体-1 凝血因子Xa |
Construction of a novel fusion protein TAP-SSL5 and evaluation of its function as anti-inflammatory and anticoagulant protein in vitro |
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| Qu Xiaolong,Hu Houyuan,Li Min,Huang Dexing,Liang Hua.Construction of a novel fusion protein TAP-SSL5 and evaluation of its function as anti-inflammatory and anticoagulant protein in vitro[J].Acta Academiae Medicinae Militaris Tertiae,2010,32(1). | |
| Authors: | Qu Xiaolong Hu Houyuan Li Min Huang Dexing Liang Hua |
| Institution: | 1Department of Cardiology;Southwest Hospital;2Department of Psychology;College of Nursing;Third Military Medical University;Chongqing;400038;China;3BakerIDI Heart & Diabetes Institute;Melbourne 8008;Australia |
| Abstract: | Objective To construct a double-effect fusion protein functioning as an anti-inflammatory and anticoagulant,which can inhibit the inflammatory response and thrombosis simultaneously in atherosclerosis lesions.Methods Firstly,staphylococcal superantigen-like protein-5(SSL5) gene sequence was cloned by PCR from Staphylococcus aureus(strain NCTC8325).Secondly,N-terminal pelB leading sequence,SSL5 gene sequence,tick anticoagulant peptide(TAP) gene sequence,with C-terminal both c-Myc and His6 tags were cloned in... |
| Keywords: | staphylococcal superantigen-like protein-5 tick anticoagulant peptide fusion protein P-selectin glycoprotein ligand-1 factor Xa |
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