Grellet F, Delcasso D, Panabieres F, Delseny M
J Mol Biol. 1986 Feb 20;187(4):495-507. doi: 10.1016/0022-2836(86)90329-3.
Two recombinant plasmids containing, respectively, three and eight tandem repeats of a 177 base-pair (bp) element from radish nuclear DNA have been isolated. These plasmids were used as probes to investigate the organization and the copy number of this element within the genome. This sequence is present in congruent to 0.6 million copies. Restriction analysis provides evidence for sequence heterogeneity and reveals the occurrence of non-overlapping subfamilies. Nine units were sequenced and found to be remarkably conserved. However, sequences in the two clones clearly belong to two distinct subgroups. Our data suggest that these sequences evolved in a concerted manner and that homogenization mechanisms such as gene conversions certainly took place. The 177 bp sequence is made from three 60 bp blocks that are derived from a common ancestor. Exchanges between the three blocks probably occurred before they became fixed as a patchwork of short sequences, the 177 bp element. This unit of 177 bp was then amplified in several steps. The presence of such a repeated sequence can be detected in other Cruciferae when hybridizations are carried out under low stringency conditions. Direct comparison with a previously published mustard satellite DNA sequence indicates a similar organization and a 75% homology. Homology was also found with shorter regions (congruent to 60 bp) of broad bean and corn satellite DNA. Finally, homology was also found with several animal alphoid sequences, suggesting that this family also occurs in the plant genomes.
已分离出两种重组质粒,它们分别含有来自萝卜核DNA的177个碱基对(bp)元件的三个和八个串联重复序列。这些质粒被用作探针来研究该元件在基因组中的组织形式和拷贝数。该序列的拷贝数约为60万份。限制性分析提供了序列异质性的证据,并揭示了非重叠亚家族的存在。对九个单元进行了测序,发现它们具有显著的保守性。然而,两个克隆中的序列明显属于两个不同的亚组。我们的数据表明,这些序列以协同方式进化,基因转换等均一化机制肯定发生过。177 bp的序列由三个60 bp的片段组成,这些片段来源于一个共同的祖先。这三个片段之间的交换可能发生在它们作为短序列的拼凑物(即177 bp元件)固定下来之前。然后,这个177 bp的单元经过几步扩增。当在低严格条件下进行杂交时,在其他十字花科植物中也能检测到这种重复序列的存在。与先前发表的芥菜卫星DNA序列直接比较表明,它们具有相似的组织形式和75%的同源性。在蚕豆和玉米卫星DNA的较短区域(约60 bp)也发现了同源性。最后,在几种动物的α卫星序列中也发现了同源性,这表明这个家族也存在于植物基因组中。