海胆基因组中长短重复序列关系的探索。
Exploration of long and short repetitive sequence relationships in the sea urchin genome.
作者信息
Eden F C, Graham D E, Davidson E H, Britten R J
出版信息
Nucleic Acids Res. 1977;4(5):1553-67. doi: 10.1093/nar/4.5.1553.
Long and short repetitive sequences of sea urchin DNA were prepared by reassociation of 2000 nucleotide long fragments to Cot 4 and digestion with the single strand specific nuclease S1. The S1 resistant duplexes were separated into long repetitive and short repetitive fractions on Agarose A50. The extent of shared sequences was studied by reassociating a labeled preparation of short repetitive DNA with an excess of unlabeled long repetitive DNA. Less than 10% of the long repetitive DNA preparation was able to reassociate with the short repetitive DNA. Thus the long and short repetitive elements appear to be principally independent sequence classes in sea urchin DNA. Precisely reassociating repetitive DNA was prepared by four successive steps of reassociation and thermal chromatography on hydroxyapatite. This fraction (3% of the genome) was reassociated by itself or with a great excess of total sea urchin DNA. The thermal stability of the products was identical in both cases (Tm=81 degrees C), indicating that precisely repeated sequences do not have many imprecise copies in sea urchin DNA.
通过将2000个核苷酸长的片段复性至Cot 4并使用单链特异性核酸酶S1消化，制备了海胆DNA的长短重复序列。用琼脂糖A50将对S1有抗性的双链体分离成长重复片段和短重复片段。通过使标记的短重复DNA制剂与过量的未标记长重复DNA复性，研究了共享序列的程度。不到10%的长重复DNA制剂能够与短重复DNA复性。因此，长重复元件和短重复元件在海胆DNA中似乎主要是独立的序列类别。通过在羟基磷灰石上进行四次连续的复性和热色谱步骤，制备了精确复性的重复DNA。该部分（占基因组的3%）自身复性或与大量过量的总海胆DNA复性。两种情况下产物的热稳定性相同（Tm = 81℃），表明在海胆DNA中精确重复的序列没有许多不精确的拷贝。
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