离子交换色谱法对大肠杆菌表达的人重组骨形态发生蛋白-7的纯化

带有BMP 7基因的大肠杆菌可以用来高量表达重组的人骨形态发生蛋白 7。升温诱导表达后 ,每升培养液大约可得到菌体湿重 3g ,其中目的蛋白约占菌体总蛋白量的 40 %。裂解离心 ,用低浓度变性剂洗涤初步纯化包涵体 ,上清中无目的蛋白损失 ,目的蛋白纯度提高到 60 %,将包涵体溶解于高浓度变性剂溶液中 ,然后在不同条件下用离子交换色谱法对变性状态下的蛋白质进行纯化 ,绝大部分杂蛋白被除去 ,目的蛋白纯度达 95 %以上 ,改变条件 ,可以减少rhBMP 7损失。并做Westernblot对目的蛋白进行特异性鉴定。

Purification of Recombinant Human Bone Morphogenetic Protein-7 on Ion Exchange Chromatography

E.coli was genetically engineered to produce recombinant human bone morphogenetic protein\|7(rhBMP\|7) in a non\|active aggregated form using a temperature\|inducible expression system.3g cell wet weight per liter of culture broth was obtained.After washing with low concentration denaturant,the protein was purified up to 60%.Further purification through ion exchange chromatography could yield rhBMP\|7 with purity exceeding 95%.Western blot was used to confirm the protein.;