A family of moderately repetitive sequences in mouse DNA.
A family of moderately repetitive sequences in mouse DNA.
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小鼠 DNA 中的一个中等重复序列家族。
DOI：
10.1093/nar/8.18.4075
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发表时间：
1980
影响因子：
14.9
通讯作者：
Schildkraut,CL
中科院分区：
文献类型：
--
作者：
Cheng,SM;Schildkraut,CL
当用限制性内切酶Eco R1酶切小鼠DNA完成时，琼脂糖凝胶电泳观察到一条1.3 kb片段的条带，约占基因组的0.5-3%。这种DNA不会在基因组中连续重复，也不是来自小鼠卫星DNA。限制性内切酶分析表明，1.3 kb片段是异质的。从1.3 kb片段中选择特异性序列，在质粒pBR322中克隆扩增。南方转移实验表明，三个独立克隆的小鼠DNA插入片段主要杂交于Eco R1 1.3 kb条带和蔗糖梯度纯化的1.3 kb片段的二次限制性内切酶切割产生的显著亚段。用Hha I （Hha I片段）从含有小鼠DNA插入的嵌合质粒中切除片段，并进行限制性内切酶和交叉杂交分析。结果发现，这三个ha I片段是不同的，尽管其中两个片段具有部分序列同源性。Cot分析表明，每个ha I片段在小鼠基因组中重复约104次。这些发现表明，在1.3 kb Eco R1波段中存在一个相关但不相同的中度重复DNA序列家族，而不是单一的均匀重复序列。
When mouse DNA is digested to completion with restriction endonuclease Eco R1, a distinct band of 1.3 kb segments comprising about 0.5–3% of the genome is observed upon agarose gel electrophoresis. This DNA is not tandemly repeated in the genome and is not derived from mouse satellite DNA. Restriction endonuclease analysis suggested that the 1.3 kb segments are heterogeneous. Specific sequences were selected from the 1.3 kb segments and amplified by cloning in plasmid pBR322. Southern transfer experiments Indicated that three separately cloned mouse DNA inserts hybridized predominantly to the Eco R1 1.3 kb band and to the conspicuous subsegments generated by secondary restriction endonuclease cleavage of the sucrose gradient purified 1.3 kb segments. Segments were also excised by Hha I (Hha I segments) from the chimeric plasmids containing mouse DNA inserts and subjected to restriction endonuclease and cross-hybridization analysis. It was found that the three Hha I segments were different, although two of them exhibited partial sequence homology. Cot analysis indicated that each of the Hha I segments are repeated about 104times in the mouse genome. These findings indicate that a family of related but non-identical, moderately repetitive DNA sequences, rather than a single homogeneous repeat, is present in the 1.3 kb Eco R1 band.