Sokolov N N, Votrin I I, Fitsner A B, Kirsanova I D, Dedov S S
Vopr Med Khim. 1980 Jul-Aug;26(4):568-72.
A new procedure is developed for isolation of highly purified preparations of restrictional endonoucleases Bam HI and Eco RI by means of fractionation with isopropyl alcohol. Restrictional endonuclease Bam HI, practically free of unspecific nucleases, was isolated after ultrasonic destruction of cells, precipitation of the restrictases with isopropanol and chromatography on DEAE cellulose. Additional chromatography on hydroxyapatite enabled to obtain the homogenous preparation of Bam HI restrictase, as shown by polyacrylamide gel disc electrophoresis. Other organic solvents (acetone, ethanol) might be also used for purification of the restrictional endonucleases.
开发了一种新方法，通过用异丙醇分级分离来分离高纯度的限制性内切酶Bam HI和Eco RI制剂。在细胞超声破碎、用异丙醇沉淀限制性酶并在DEAE纤维素上进行色谱分离后，分离出几乎不含非特异性核酸酶的限制性内切酶Bam HI。如聚丙烯酰胺凝胶圆盘电泳所示，在羟基磷灰石上进一步色谱分离能够获得Bam HI限制性酶的纯制剂。其他有机溶剂（丙酮、乙醇）也可用于纯化限制性内切酶。
