两步亲和层析纯化限制性核酸 内切酶Bsp 63Ⅰ
Purification of restriction endonuclease Bsp63Ⅰ by two step affinity chromatography
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摘要: 以Bacillus sphaericus 63为材料， 采用DNA-Sepharose 4B 和Cibacron Blue F3GA-Sepharose 4B两步亲和层析， 分离纯化得到电泳均一纯限制性核酸内切酶Bsp 63 Ⅰ. 酶的得率约为130单位/g湿菌， 比活力高达61400单位/mg蛋白. 还报道了DNA-Speharose 4B两种亲和吸附剂制作方法的改进.Abstract: Restriction endonuclease Bsp63Ⅰ has been isolated and purified by affinity chromatography on self- made DNA Sepharose4B and Cibacron Blue F3GA Sepharose4B from Bacillus sphaericus 63. The purified enzyme was found to be homogeneous as judged by polyacrylamide gel electrophoresis. The specific activity of the enzyme is greater than 61400 units per mg protein and the yield of the purified enzyme is greater than 130 units per gram wet cells.
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