Lobanenkov V V, Mironov N M, Kuprianova E I, Shapot V S
Biokhimiia. 1985 Jul;50(7):1132-40.
Isolated cell nuclei were incubated with nucleases followed by extraction of chromatin with a low salt buffer. With an increase of nuclear chromatin degradation with DNAse I or micrococcal nuclease, solubilization of deoxyribonucleoprotein (DNP) by a low salt buffer increases, reaching a maximum upon hydrolysis with 2-4% nuclear DNA and then decreases appreciably after extensive treatment with nucleases. Soluble fragmented chromatin aggregates in the course of treatment with DNAase. I. Addition to gel chromatin preparations of exogenous products of nuclease treatment of isolated nuclei leads to its aggregation. Pretreatment of nuclear chromatin with RNAase prevents solubilization of DNP by low ionic strength solutions. Some experimental data obtained with the use of severe nuclease treatment are discussed; for a correct interpretation of these data the aggregation of fragmented chromatin by products of its nuclease degradation should be taken into consideration.
将分离的细胞核与核酸酶一起孵育,然后用低盐缓冲液提取染色质。随着DNA酶I或微球菌核酸酶对核染色质降解的增加,低盐缓冲液对脱氧核糖核蛋白(DNP)的溶解作用增强,在用2-4%的核DNA水解时达到最大值,然后在核酸酶大量处理后明显下降。在DNA酶I处理过程中,可溶性片段化染色质会聚集。向凝胶染色质制剂中添加经核酸酶处理的分离细胞核的外源产物会导致其聚集。用RNA酶对核染色质进行预处理可防止低离子强度溶液溶解DNP。讨论了使用剧烈核酸酶处理获得的一些实验数据;为了正确解释这些数据,应考虑染色质片段被其核酸酶降解产物聚集的情况。