Tseng B Y, Goulian M
Cell. 1977 Oct;12(2):483-9. doi: 10.1016/0092-8674(77)90124-6.
A short RNA covalently associated with nascent DNA has been isolated after synthesis in vitro with labeled ribonucleaside triphosphates and the removal of DNA by DNAase digestion. The RNA migrates in polyacrylamide gels or chromatographs on DEAE-Sephadex columns as a relatively discrete oligonucleotide 8-11 nucleotides in length. The RNA is associated primarily with nascent DNA with stoichiometry of approximately one per DNA chain. The RNA has a triphosphate group at the 5' end and 2 or 3 deoxynucleotide residues at the 3' end that are not removed by DNAase. These results further support a role for the RNA as an initiator of discontinuous DNA synthesis. Examination of sequences present at the 3' end of the RNA using RNAase to effect transfer of 32PO4 from 32P-labeled DNA to covalently attached RNA indicates that a diverse, rather than unique, set of sequences are present in the RNA.
在体外使用标记的核糖核苷三磷酸进行合成并通过DNA酶消化去除DNA后，已分离出一种与新生DNA共价结合的短RNA。该RNA在聚丙烯酰胺凝胶中迁移，或在DEAE-葡聚糖凝胶柱上进行色谱分析，呈现为长度约为8至11个核苷酸的相对离散的寡核苷酸。该RNA主要与新生DNA结合，化学计量比约为每条DNA链一个。该RNA在5'端有一个三磷酸基团，在3'端有2或3个脱氧核苷酸残基，这些残基不会被DNA酶去除。这些结果进一步支持了RNA作为不连续DNA合成起始物的作用。使用RNA酶将32P标记的DNA中的32PO4转移到共价连接的RNA上，以检查RNA 3'端存在的序列，结果表明RNA中存在的是一组多样而非独特的序列。
