CNTF工程菌培养及产物纯化

Culture of Ciliary Nerves Trophic Factor (CNTF) Engineering Bacterial Strain and Purification of Expressed Product

目的 建立ＣＮＩＦ生产工艺，获得高产量的ＣＮＴＦ活性蛋白质。方法 通过摇瓶和发酵罐培养研究 了培养基成分、ｐＨ、诱导时间对工程菌 ｐＢＶ２２０－ＣＮＴＦ／ＤＨ５Ａα生成和表达的影响，并对表达产物进行纯化和生物学活 性检测。结果 应用正交试验确定了培养基成分配比为酵母粉１０ｇ／Ｌ，葡萄糖８ｇ／Ｌ，ＭｇＳＯ４０．５ｇ／Ｌ，磷酸盐０．０５ｍｏｌ／Ｌ， ｐＨ７．４时最有利于工程菌的生长及ＣＮＩＴ的表达。ＣＮＴＴ纯化后纯度达９５％以上，并具有较高的生物学活性。结论 为ＣＮＴＦ的结构和功能的研究及临床应用打下基础。

Objective To obtain a large amount of active ciliary nerves trophic factor (CNTF) protein by gene engineering technique. Methods The influences of medium component, pH and induction time on the growth and expression of engineering bacterial strain pBV22O - CNTFIDH5α were studied. The expressed product was purified, and the biological activity of it was detected. Results The optimal component of medium was defined as yeast powder( 10g/L) , glucose(8g/L) and MgSO4 (0.5g/L) ,and the optimal phosphate concentration and pH value were 0. 05mol/L and 7.4 respectively. The expressed CNTF showed high biological activity and reached a purity of above 95% after being purified. Conclusion This paper laid a foundation of study on the structure, function and clinical application of CNTF.