用限制性核酸酶制备可溶性染色质和特定染色质组分。

Igo-Kemenes T, Greil W, Zachau H G

Nucleic Acids Res. 1977 Oct;4(10):3387-400. doi: 10.1093/nar/4.10.3387.

By digestion of rat liver nuclei with EndoR HaeIII, EndoR EcoRI, and EndoR Bam and subsequent lysis of the nuclei approx. 90%, 40%, and 45%, respectively, of the chromatin were solubilized. The plateau values of solubilization are in agreement with a model in which the chromatin strands are crosslinked and/or attached to a supporting structure. The distribution of DNA lengths in the soluble and insoluble chromatin fractions were determined. According to digestion experiments with restriction nucleases rat liver DNA contains highly repetitive sequences, some of which are arranged in tandem repeats of 95 and 380 nucleotide pairs, respectively. With EndoR EcoRI chromatin containing the repetitive RNA was preferentially solubilized and, by subsequent sucrose gradient centrifugation, purified to about 90%. The useful properties of chromatin prepared by the specific action of restriction nucleases are discussed.

用内切酶HaeIII、内切酶EcoRI和内切酶Bam消化大鼠肝细胞核，随后裂解细胞核，分别约90%、40%和45%的染色质被溶解。溶解的平稳值与染色质链交联和/或附着于支持结构的模型一致。测定了可溶性和不溶性染色质组分中DNA长度的分布。根据用限制性核酸酶进行的消化实验，大鼠肝DNA含有高度重复序列，其中一些分别排列成95和380个核苷酸对的串联重复序列。用内切酶EcoRI处理时，含有重复RNA的染色质优先被溶解，随后通过蔗糖梯度离心，纯化至约90%。讨论了通过限制性核酸酶的特异性作用制备的染色质的有用特性。