Novel classes of mouse repeated DNAs.
Novel classes of mouse repeated DNAs.
复制标题
新型小鼠重复 DNA 类别。
DOI：
10.1093/nar/8.15.3247
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发表时间：
1980
影响因子：
14.9
通讯作者：
Manuelidis,L
中科院分区：
文献类型：
--
作者：
Manuelidis,L
用Eco RII （Bst NI）切割的小鼠DMA在溴化乙酯染色的琼脂糖凝胶中显示出1.5和1.7 kb的显著限制性带。这些构成了小鼠重复DNA的新亚群。序列Hoechst 33258-CsCl梯度分离小鼠DNA，得到更多的富含GC的主带DNA和富含AT的卫星DNA，表明这两个片段都与富含GC的主带DNA共增殖。在纯化的卫星制剂中未检测到它们。这些限制性内切带合计构成主带dna≥0.2%。将32P标记的卫星DNA与Eco RII限制性小鼠DNA进行杂交，结果显示只有较小的卫星限制性片段出现阳性杂交，这表明卫星DNA与1.5或1.7 kb片段几乎没有同源性。从凝胶中分离1.5 kb和1.7 kb的片段，用刻痕翻译标记32p。利用一系列限制性内切酶，这两个片段显示出不同的切割模式。过滤杂交证实这两个片段是不同的子集，因为它们彼此没有交叉杂交。它们也没有与溴化乙酯染色凝胶中注意到的其他次要的重复非卫星DNA带杂交。它们都不能与32p激酶标记的18S和28S RNA杂交，因此不能被分配到核糖体基因上。从雄性和雌性小鼠中分离的DNA显示1.5 kb和1.7 kb片段的数量相当。因此，两者都不是Y染色体特异性的。从限制模式和初步的染色体杂交研究来看，这些片段被认为是“穿插”的重复序列，而不是很长的串联（卫星状）着丝粒排列。这些重复序列亚群之间的关系，它们的进化和详细组织，以及它们在不同小鼠物种中的代表性，仍有待确定。
Mouse DMA cleaved with Eco RII (Bst NI) displays two prominent restriction bands of 1.5 and 1.7 kb in agarose gels stained with ethtdlum bromide. These constitute novel subsets of repeated DNA in the mouse. Sequential Hoechst 33258-CsCl gradient fractionetion of mouse DNA, yielding more GC rich main band DNA, and AT rich satellite DNA, revealed that both these fragments copurified with GC rich main band DNA. They were not detected in purified satellite preparations. Together these restriction bands constltuted ≥0.2% of main band DNAs. Hybridization of 32P labelled satellite DNA to blots of Eco RII restricted mouse DNA showed positive hybridization only to smaller satellite restriction fragments, indicating satellite DNA had little or no homology with either the 1.5 or 1.7 kb fragments.The 1.5 and 1.7 kb fragments were isolated from gels and labelled with32P by nick translation. Using a series of restriction endonucleases each of these two fragments showed different cleavage patterns. Filter hybridization confirmed that these two fragments were distinct subsets as they did not cross hybridize with each other. They also did not hybridize to other more minor repeated non-satellite DNA bands noted in ethidlum bromide stained gels. Neither of them could be assigned to ribosomal genes as they did not hybridize to32p kinase labelled 18S and 28S RNA. Isolation of DNA from male and female mice showed comparable amounts of both the 1.5 and 1.7 kb fragments. Thus neither was Y chromosome specific. From restriction patterns, and preliminary chromosome hybridization studies, these fragments are thought to represent “interspersed” repeated sequences rather than very long tandem (satellite like) centromeric arrays.The relationship between these repeated sequence subsets, their evolution and detailed organization, and their representation in different mouse species, remain to be determined.