Choo K H, Brown R, Webb G, Craig I W, Filby R G
Murdoch Institute for Research into Birth Defects, Royal Children's Hospital, Parkville, Victoria, Australia.
DNA. 1987 Aug;6(4):297-305. doi: 10.1089/dna.1987.6.297.
We characterized a recombinant clone E7 containing a 1.6-kb Eco RI insert of human alpha satellite DNA (alpha DNA) which hybridized in situ predominantly to the centromere of chromosome 17. Three thousand copies of this sequence were detected on chromosome 17, although a lesser number of copies were also found on the centromeres of chromosomes 11, X, and the other human chromosomes, except Y. In the human genome, sequences homologous to E7 were organized principally as five major polymorphic (Pst I) forms of tandem alpha DNA repeats with molecular weights between 2.0 and 2.7 kb. We We studied the higher-order organization of these major forms using a series of 12 cosmid clones. Close linkage of the different polymorphic forms was demonstrated, with no two cosmids showing an identical linkage pattern. Six of the cosmid clones carried a considerable amount (20-25%) of nonhomologous (non-alpha) DNA, indicating that the repeat arrays are relatively frequently interrupted by other genomic DNA. In none of the cosmid inserts were the repeat arrays bound on both sides by non-alpha DNA, suggesting that short arrays are not common. However, some of the intervening non-alpha DNA sequences were relatively short, and vary in size from 6 to 24 kb. Our results suggest an irregular and complex pattern of organization of alpha DNA in the human genome.
我们鉴定了一个重组克隆E7,它含有一段1.6kb的人类α卫星DNA(αDNA)的Eco RI插入片段,该片段原位杂交主要定位于17号染色体的着丝粒。在17号染色体上检测到3000个该序列的拷贝,不过在11号染色体、X染色体以及除Y染色体外的其他人类染色体的着丝粒上也发现了较少数量的拷贝。在人类基因组中,与E7同源的序列主要组织为5种主要的多态性(Pst I)形式的串联αDNA重复序列,分子量在2.0至2.7kb之间。我们使用一系列12个黏粒克隆研究了这些主要形式的高级组织。结果表明不同多态性形式紧密连锁,没有两个黏粒显示出相同的连锁模式。6个黏粒克隆携带了相当数量(20 - 25%)的非同源(非α)DNA,这表明重复阵列相对频繁地被其他基因组DNA打断。在任何一个黏粒插入片段中,重复阵列两侧都没有被非αDNA所界定,这表明短阵列并不常见。然而,一些插入的非αDNA序列相对较短,大小在6至24kb之间。我们的结果表明人类基因组中αDNA的组织模式不规则且复杂。