人TPON端结构域在大肠杆菌中的表达、纯化及活性分析  

Exppression and Purification the N terminal Domain of Human TPO in E.coli

作　　者：刘丽[1] 田生礼[1] 王艳华 孟岩[1] 芦兴武 谢宝树[1] 

机构地区：[1]空军总医院空军临床分子生物学研究中心

出　　处：《生物化学杂志》1997年第6期666-671,共6页

基　　金：全军"九五"医药卫生科研基金

摘　　要：在利用反转录－ＰＣＲ从人胎肝中获得编码人血小板生成素（ｈＴＰＯ）全长ｃＤ－ＮＡ的基础上，综合ＴＰＯ结构与功能的研究信息，在大肠杆菌中表达了成熟肽Ｎ端结构域．目的蛋白在菌体内以包涵体形式存在，表达量约占菌体总蛋白的３０％；包涵体经变性、复性、凝胶过滤、离子交换层析等步骤处理后，所得产物给Ｂａｂｌ／ｃ小鼠腹腔连续注射８ｄ，第９ｄ摘眼球采血，计数血小板的数量．结果表明，ＴＰＯＮ端结构域具有明显促进血小板生成的作用．The full length cDNA of human thrombopoitin (hTPO) from fetal liver had been cloned previously by RT PCR. Refering to the new achievements of TPO structure and function research, a truncated N terminal domain of mature hTPO was expressed in E. coli .Amount of TPO expressed was 30% of total bacterial protein in the form of inclusion. After denaturation, renaturation, gel and ion exchange chromatography,the truncated hTPO was purified.It was administered to Babl/c mice peritoneally for each day 8 days, and the number of periperal platelets were counted on the 9th day. It was then confirmed that TPO N terminal domain could stimulate and improve pletelet production.

关 键 词：血小板生成素 遗传工程 药物 N端结构域 研制 

分 类 号：R392－33[医药卫生—免疫学] R394.8[医药卫生—基础医学]