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哺乳动物中与卫星DNA相邻的低拷贝数序列的分离。

Isolation of low-copy-number sequences that neighbor satellite DNA in mammals.

作者信息

Maresca A, Thayer R E, Guenet C, Singer M F

出版信息

Gene. 1986;50(1-3):299-311. doi: 10.1016/0378-1119(86)90334-3.

Abstract

To investigate the role of satellite DNA in eukaryotic genomes, we isolated from an African green monkey (Cercopithecus aethiops) genomic library cloned segments containing the previously described deca-satellite linked to low-copy-number genomic sequences. Three such clones were obtained. The low-copy-number sequences in the three clones do not cross-hybridize suggesting that they derive from different genomic loci. The structure of one of the clones, lambda MkA, is described in detail. Subcloned segments containing the low-copy-number sequences from lambda MkA anneal to monkey, human and mouse genomic DNA. The subcloned probes were used to select clones containing homologous sequences from a second, independent monkey library as well as from human and mouse genomic libraries. Several of the newly isolated monkey clones hybridized to probes containing the species-specific deca- and alpha-satellites, confirming the genomic association of the low-copy-number sequence in lambda MkA with satellite DNA. Moreover, several of the human and mouse clones hybridized to species-specific human and mouse satellite DNAs, respectively. These experiments indicate that the low-copy-number sequence in lambda MkA and its association with satellite DNA is conserved in primates and rodents.

摘要

为了研究卫星DNA在真核生物基因组中的作用,我们从非洲绿猴(Cercopithecus aethiops)基因组文库中分离出了含有先前描述的与低拷贝数基因组序列相连的十聚体卫星的克隆片段。获得了三个这样的克隆。这三个克隆中的低拷贝数序列不发生交叉杂交,表明它们来自不同的基因组位点。详细描述了其中一个克隆λMkA的结构。从λMkA中含有低拷贝数序列的亚克隆片段与猴、人和小鼠的基因组DNA退火。亚克隆探针用于从第二个独立的猴文库以及人和小鼠基因组文库中筛选含有同源序列的克隆。几个新分离的猴克隆与含有物种特异性十聚体和α卫星的探针杂交,证实了λMkA中低拷贝数序列与卫星DNA的基因组关联。此外,几个人和小鼠克隆分别与物种特异性的人和小鼠卫星DNA杂交。这些实验表明,λMkA中的低拷贝数序列及其与卫星DNA的关联在灵长类动物和啮齿动物中是保守的。

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