Razin S V, Mant'eva V L, Georgiev G P
Mol Biol (Mosk). 1980;14(1):223-33.
DNA fractions enriched in sequences located closely to the sites of attachment of DNA loops to the axial protein structures of interphase nuclei ad metaphase chromosomes were isolated with the aid of two techniques. The method involving treatment with EcorI endonuclease has been described earlier. The new method is based on the mild treatment of the preparations with micrococcal nuclease. The latter method allows one to obtain the fraction of attached DNA of different length and to compare their properties. The average distance between two attachment sites was measured. It is equal to about 6.10(4) base pairs in the both interphase nuclei and metaphase chromosomes. The attached DNA obtained with the aid of micrococcal nuclease treatment is not enriched in satellite but contains a high amount of middle repetitive sequences renaturing in the Cot interval from 10(-2) to 10(0). According to renaturation properties and distribution in CsCl density gradient the fractions of attached DNA of similar size from metaphase chromosomes and from interphase nuclei are virtually identical. The data obtained indicate that the DNA sequences involved in the attachment of DNA loops to the matrix of interphase nucleus are the same as in the case of metaphase chromosomes.
借助两种技术分离出了富含与DNA环附着于间期细胞核和中期染色体轴向蛋白质结构位点紧密相邻序列的DNA片段。涉及用EcorI核酸内切酶处理的方法先前已作过描述。新方法基于用微球菌核酸酶对制剂进行温和处理。后一种方法能够获得不同长度的附着DNA片段,并比较它们的特性。测量了两个附着位点之间的平均距离。在间期细胞核和中期染色体中,该距离均约为6×10⁴个碱基对。借助微球菌核酸酶处理获得的附着DNA在卫星DNA中并不富集,但含有大量在Cot区间从10⁻²到10⁰复性的中等重复序列。根据复性特性以及在氯化铯密度梯度中的分布,来自中期染色体和间期细胞核的大小相似的附着DNA片段实际上是相同的。所获得的数据表明,参与DNA环附着于间期细胞核基质的DNA序列与中期染色体的情况相同。