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EcoRI 限制性内切酶的溶解性及其从高产菌株中的纯化。

Solubility of the EcoRI restriction endonuclease and its purification from an over-producing strain.

作者信息

Luke P A, Halford S E

出版信息

Gene. 1985;37(1-3):241-6. doi: 10.1016/0378-1119(85)90278-1.

Abstract

The solubility of the EcoRI restriction endonuclease was measured in solutions of varying NaCl concentrations, at different temperatures and in the presence of DNA. The precipitation of the protein was enhanced by low NaCl concentrations, by elevated temperatures, and by the addition of DNA. These observations are discussed in relationship to the interaction of this protein with DNA. The purification of the EcoRI restriction enzyme from a strain of Escherichia coli that over-produces this enzyme was hampered by the insolubility of the protein, and hence the purification procedure was modified to optimize the recovery of active enzyme.

摘要

在不同NaCl浓度、不同温度以及存在DNA的溶液中,测定了EcoRI限制性内切核酸酶的溶解度。低NaCl浓度、升高温度以及添加DNA都会增强蛋白质的沉淀。结合该蛋白质与DNA的相互作用对这些观察结果进行了讨论。从一株过量产生该酶的大肠杆菌菌株中纯化EcoRI限制酶时,由于蛋白质的不溶性而受到阻碍,因此对纯化程序进行了修改,以优化活性酶的回收率。

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