Highly repeated DNA families in the rat.
Highly repeated DNA families in the rat.
复制标题
大鼠体内高度重复的 DNA 家族。
DOI：
10.1016/s0021-9258(18)90989-0
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
A. Furano
中科院分区：
文献类型：
--
作者：
F. Witney;A. Furano
我们已经分析了重复的DNA部分的大鼠通过表征约500个重复的DNA含有克隆使用杂交到各种啮齿动物的核酸。为了便于这种分析，我们设计了一种方法，通过直接从细菌克隆的菌落中印迹，将克隆的DNA转移到硝酸纤维素纸上。除了鉴定潜在感兴趣的重复序列（例如，以组织特异性方式转录的那些序列，或在非大鼠基因组中高度保守的那些序列）之外，我们发现，与通过大鼠DNA的重新缔合所揭示的相反（例如，Pearson，W. R.，吴，J.R.，和Bonner，J.（1978）Biochemistry 17，51-59），大鼠基因组含有许多不同的高度重复（大于50，000个拷贝）序列。我们通过与不同核酸的杂交以及DNA序列测定来区分不同的高度重复序列。这些高度重复的序列有三个共同的特征，使它们与10万个成员的大鼠卫星I家族区别开来：（i）它们在克隆的重复DNA文库中的回收率低于它们在大鼠基因组中拷贝数的预期;（ii）它们的拷贝数再退火异常缓慢，即使它们没有显著的差异;和（iii）它们在一种或多种大鼠组织中转录。这些研究结果的组织在大鼠基因组中的重复序列的影响进行了讨论。
We have analyzed the repeated DNA fraction of the rat by characterizing approximately 500 repeat DNA-containing clones using hybridization to a variety of rodent nucleic acids. To facilitate this analysis we devised a method whereby the cloned DNA is transferred to nitrocellulose paper by blotting directly out of colonies of the bacterial clones. In addition to identifying repeated sequences of potential interest (e.g. those transcribed in a tissue-specific manner, or those that are highly conserved in non-rat genomes), we found that, in contrast to what is revealed by the reassociation of rat DNA (e.g. Pearson, W. R., Wu, J. R., and Bonner, J. (1978) Biochemistry 17, 51-59), the rat genome contains a number of different highly repeated (greater than 50,000 copies) sequences. We distinguished the different highly repeated sequences both by their hybridization to different nucleic acids as well as by DNA sequence determination. The highly repeated sequences shared three characteristics that distinguished each of them from the 100,000-member rat satellite I family: (i) they were recovered less often in the cloned repeat DNA library than expected from their copy number in the rat genome; (ii) they reannealed abnormally slowly for their copy number even though they are not significantly divergent; and (iii) they are transcribed in one or more rat tissues. The implications of these findings for the organization of repeated sequences in the rat genome are discussed.