重组人类肥胖抑素在大肠杆菌中的大规模纯化

Large -scale purification of recombinant human leptin from recombinant E.Coli

目的 探讨产率较高 ,高度纯化大肠杆菌表达重组肥胖抑素的方法 ,深入研究肥胖抑素的分子结构 ,生物活性及临床应用 ,对在大肠杆菌中表达的重组人类肥胖抑素进行纯化。方法 温度诱导表达携有人类肥胖基因 (ob)片段的大肠杆菌重组子 ,采用酸沉淀结合凝胶过滤层析的方法从包涵体中纯化目的蛋白。结果 1周内从 3L培养液 (10g)湿菌体中可获得 70mg高度纯化的重组人类肥胖抑素。结论 在短期内成功获得了高产高纯化的重组人类肥胖抑素 ,为肥胖抑素的产业化及临床应用奠定了基础。

Objective To purify recombinant human leptin from E.Coli for investigating its characteristics, biological activity and clinical application. Methods The E.Coli DH5 α cells transfected with recombinant pBV220-OB, a human obese gene. The expressed protein found in the inclusion bodies was purified by using acid precipitation and chromatography. Results Seventy milligrams of leptin with a high purity was prepared from 3 liter E.Coli culture media (10 grams of cells) within a week. Conclusion Large quantity of highly purified human leptin could be succesfully and rapidly prepared; it will hasten investigation of its production and clinical application.