Gorodetskii S I, Adler V V, Vasil'eva I M, Zlatopol'skii A T, Mazin A L
Mol Biol (Mosk). 1976 Jul-Aug;10(4):609-19.
A procedure for the isolation and purification of a specific hybrid between rat 28S and 18S ribosomal RNA's and nucleolar DNA is described. The method employed includes the following steps: 1) isolation of the nucleolar DNA, 2) hybridization of [14C]rRNA with the nucleolar DNA, and 3) isolation and purification of the rRNA-DNA hybrid complex by chromatography on hydroxylapatite and centrifugation in a CsCl density gradient. In the isolated hybrid complex the RNA:DNA ratio is close to 1:1, and the degree of enrichment of the DNA by the rRNA cistrons is about 1500 times. The hybrid obtained has a sedimentation constant on the order of 20S, is resistant to the action of pancreatic RNase and RNase T1 and sheep brain DNase, and is characterized by high thermostability. Acording to the physicochemical tests used, the rRNA-DNA hybrid complex is a double-stranded poly-nucleotide with an ordered secondary structure.
本文描述了一种分离和纯化大鼠28S与18S核糖体RNA及核仁DNA之间特定杂交体的方法。所采用的方法包括以下步骤：1）核仁DNA的分离；2）[14C]rRNA与核仁DNA的杂交；3）通过羟基磷灰石柱层析和CsCl密度梯度离心对rRNA-DNA杂交复合体进行分离和纯化。在分离得到的杂交复合体中，RNA与DNA的比例接近1:1，rRNA顺反子对DNA的富集程度约为1500倍。所获得的杂交体沉降常数约为20S，对胰核糖核酸酶、核糖核酸酶T1和羊脑脱氧核糖核酸酶的作用具有抗性，并且具有高热稳定性。根据所采用的物理化学测试，rRNA-DNA杂交复合体是一种具有有序二级结构的双链多核苷酸。
