Department of Biotechnology, Osaka University, Suita, Japan.
Mol Biotechnol. 2010 Feb;44(2):148-51. doi: 10.1007/s12033-009-9223-3.
We report a simplified alternative protocol for purification of recombinant linker histone H1 under non-denaturing conditions. This method takes advantage of the strong affinity of H1 to DNA and comprises nucleoprotein complex extraction from the lysate of bacterial cells overexpressing the protein, followed by two ion-exchange purification steps. The purity of the protein was at least 95%; the purified H1 was tested for nucleosome binding and was successfully fluorescently labeled for further studies.
我们报告了一种在非变性条件下纯化重组连接组蛋白 H1 的简化替代方案。该方法利用 H1 与 DNA 的强亲和力，包括从表达该蛋白的细菌细胞裂解物中提取核蛋白复合物，然后进行两步离子交换纯化。蛋白质的纯度至少为 95%；纯化的 H1 进行了核小体结合测试，并成功地进行了荧光标记，以进行进一步研究。
