Cavalieri L F, Carroll E
Proc Natl Acad Sci U S A. 1970 Oct;67(2):807-12. doi: 10.1073/pnas.67.2.807.
A new procedure is described for purifying proteins that specifically bind to DNA. DNA is entrapped in polyacrylamide gel particles which can then be used in standard column chromatographic procedures. The method was developed using Escherichia coli DNA polymerase as the test material. The crude enzyme was applied at low ionic strength and eluted at high ionic strength with a 200-fold increase in specific activity on a single passage through the column. The method is versatile and simple and is not restricted to DNA-protein systems. Any macromolecule can be entrapped in the gel particles; these can interact with other large or small molecules in the liquid phase. The gel is stable at elevated temperatures and can therefore be used in hybridization experiments.
本文描述了一种纯化与DNA特异性结合的蛋白质的新方法。DNA被包埋在聚丙烯酰胺凝胶颗粒中,然后可用于标准柱色谱程序。该方法是使用大肠杆菌DNA聚合酶作为测试材料开发的。粗酶在低离子强度下上样,在高离子强度下洗脱,单次通过柱子时比活性提高了200倍。该方法通用且简单,不限于DNA-蛋白质系统。任何大分子都可以包埋在凝胶颗粒中;这些颗粒可以与液相中的其他大分子或小分子相互作用。该凝胶在高温下稳定,因此可用于杂交实验。