核小体的提取纯化及鉴定

Isolation, purification and identification of nucleosomes from chicken erythrocytes

目的 探索一种操作简便、实用的核小体提取方法。方法 取无菌新鲜鸡血 ,提取细胞核 ,用葡萄球菌核酶消化后裂解、离心 ,获得上清组分用连续性密度梯度蔗糖进行分离 ,收集分子量 3 5 0bp左右的组分。结果 2 %琼脂糖电泳显示该组分含有长度为 15 0bp的双链DNA ,15 %SDS PAGE电泳显示该组分含有组蛋白H2A、H2B、H3和H4;本实验分离的核小体在 0℃条件下存放 4周未见DNA成分和组蛋白成分发生解聚。结论 该方法可以正确的分离出核小体 ,且具有良好的稳定性 ,为深入研究核小体在SLE自身抗体产生中的作用打下基础。

Objective To establish a simple and practical method for isolating and purifying nucleosomes. Methods Nuclei were isolated from chicken erythrocytes, and then digested with staphylococcal nuclease. After centrifugation, the supernatant of digestion was separated and centrifugated on sucrose gradients. Results Nucleosomes with good stability were isolated properly by gradient centrifugation. Conclusion This method for the isolation and purification of nucleosomes is simple and effective, which might contribute to the further researches of the roles of nucleosomes in the pathogenesis of systemic lupus erythematosus.