Kipling D, Ackford H E, Taylor B A, Cooke H J
MRC Human Genetics Unit, Western General Hospital, Edinburg, United Kingdom.
Genomics. 1991 Oct;11(2):235-41. doi: 10.1016/0888-7543(91)90128-2.
As an adjunct to attempts to define functionally important sequences at human centromeres, we have undertaken a long-range physical analysis of these regions in the mouse. Mouse centromeres are usually situated very close to the chromosome ends and are closely associated with minor satellite sequences on the basis of cytological observations. Using pulsed-field gel electrophoresis we find that this satellite DNA is arranged as tandem arrays, predominantly uninterrupted by nonsatellite sequences. These arrays can be released largely intact by digestion with a range of enzymes that generally cleave frequently in non-satellite DNA. The restriction fragments carrying these arrays are polymorphic in size between inbred strains and provide direct markers for mouse centromeres. To illustrate the possible use of these polymorphic markers we have mapped a 1.3-Mb PvuII variant in a set of RI strains to the centromere of Chromosome 7. The minor satellite arrays are very close to the centromeric telomere and physical linkage with terminal repeat sequences can readily be detected, placing many minor satellite arrays on terminal restriction fragments smaller than 1 Mb. The apparent lack of any sizable amount of nonsatellite DNA between the minor satellite and the terminal repeat arrays indicates that many mouse chromosomes are truly telocentric.
作为确定人类着丝粒功能重要序列的辅助手段,我们对小鼠的这些区域进行了长期的物理分析。根据细胞学观察,小鼠着丝粒通常位于非常靠近染色体末端的位置,并且与小卫星序列密切相关。使用脉冲场凝胶电泳,我们发现这种卫星DNA以串联阵列的形式排列,主要不被非卫星序列打断。通过用一系列通常在非卫星DNA中频繁切割的酶进行消化,这些阵列可以在很大程度上完整地释放出来。携带这些阵列的限制性片段在近交系之间大小具有多态性,并为小鼠着丝粒提供了直接标记。为了说明这些多态性标记的可能用途,我们在一组RI品系中将一个1.3-Mb的PvuII变体定位到了7号染色体的着丝粒上。小卫星阵列非常靠近着丝粒端粒,并且可以很容易地检测到与末端重复序列的物理连锁,许多小卫星阵列位于小于1 Mb的末端限制性片段上。小卫星与末端重复阵列之间明显缺乏任何可观数量的非卫星DNA,这表明许多小鼠染色体实际上是端着丝粒染色体。