人TPON端结构域在大肠杆菌中的表达、纯化及活性分析

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．

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

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.