细胞凋亡检测蛋白AnnexinV的制备纯化和初步评价  

Preparation and in Vitro Evaluation of Human Recombinant Annexin V Using for Apoptosis

作　　者：陈大明[1] 谢虹 祁本忠[1] 张颖梅 贾兵[3] 杨红伟[1] 罗志福[1] 张锦荣[1] 金小海[1] 王凡[3] 马大龙[2] 

机构地区：[1]中国原子能科学研究院同位素研究所,北京102413 [2]北京大学人类疾病基因研究中心,北京100083 [3]北京大学医学同位素研究中心,北京100083

出　　处：《原子能科学技术》2004年第z1期182-187,共6页Atomic Energy Science and Technology

摘　　要：文章利用基因工程技术在大肠杆菌中高效表达了人重组AnnexinV,建立了批量获得AnnexinV工程菌诱导表达的最佳温度、时间和包涵体的操作程序。通过SDS PAGE分析和得到的AnnexinV标记上FITC后能对地塞米松引起Balb/c小鼠胸腺单细胞产生的凋亡进行有效检测的实验表明:经过离子层析纯化后得到了高纯度、有生物活性的人重组AnnexinV纯品。细胞结合分析的结果显示:AnnexinV的KD值为8.53nmol/L,RT为8.79nmol/L。Human recombinant Annexin V was produced by expression in E coli with high efficiency through genetic engineering. The technique procedure concerned in the temperature and time of vector expression, the basic routine and purification of proteins was established in order to obtain a large quantity of Annexin V. The results of SDS-PAGE analysis and the apoptosis detection of single cell of thymocytes of Balb/c mice (using) FITC-Annexin V caused by dexamethasone with availableness show that the mature Annexin V with high purity and biologic activity is obtained by ion exchange chromatography. The results of cell binding assay show that its KD is 8.53 nmol/mL

关 键 词：细胞凋亡 ANNEXIN V 诱导表达和纯化 FITG-Annexin V 细胞结合分析 

分 类 号：R34[医药卫生—基础医学]