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质粒DNA与蛋白质的松弛复合体。I. 质粒ColE1和ColE2松弛复合体中蛋白质与DNA的链特异性结合

Relaxation complexes of plasmid DNA and protein. I. Strand-specific association of protein and DNA in the relaxed complexes of plasmids ColE1 and ColE2.

作者信息

Blair D G, Helinski D R

出版信息

J Biol Chem. 1975 Nov 25;250(22):8785-9.

PMID:1102543
Abstract

The ColE1 and ColE2 relaxation complexes of supercoiled DNA and protein were purified from Escherichia coli cells. Protein remains associated with the open circular DNA of these complexes after induction of relaxation with sodium dodecyl sulfate. The protein is associated specifically with the strand that possesses a site-specific break in the Co1E1 and ColE2 relaxed complexes. This protein remains associated with the DNA after centrifugation of the relaxed complex in a neutral or alkaline (pH 12.5) cesium chloride gradient or treatment with 8 M urea, 2 M NaSCN, 2M LiCl, 0.2 M sodium acetate, pH 4.6, and 70% formamide at 60 degrees.

摘要

从大肠杆菌细胞中纯化出超螺旋DNA与蛋白质的ColE1和ColE2松弛复合体。在用十二烷基硫酸钠诱导松弛后,蛋白质仍与这些复合体的开环DNA结合。该蛋白质与在ColE1和ColE2松弛复合体中具有位点特异性断裂的链特异性结合。在用中性或碱性(pH 12.5)氯化铯梯度对松弛复合体进行离心,或用8M尿素、2M硫氰酸钠、2M氯化锂、0.2M乙酸钠(pH 4.6)和60℃下的70%甲酰胺处理后,该蛋白质仍与DNA结合。

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