Cardone D E, Feliciello I, Marotta M, Rosati C, Chinali G
Dipartimento di Medicina Clinica e Sperimentale, Facoltà di Medicina e Chirurgia, Università di Napoli, Federico II, Italy.
Genome. 1997 Oct;40(5):774-81. doi: 10.1139/g97-800.
Digestion of Rana graeca italica DNA with Asp 718I produces highly repetitive fragments of 281 and 385 bp that were cloned and sequenced. The shorter fragment corresponds to the unit repeat (RgiS1b) of a satellite DNA. The longer fragment was found to be part of a 494-bp repeat of another satellite DNA (RgiS1a) that was cloned intact as an EcoRV fragment. RgiS1b is 97% homologous to RgiS1a, from which it seems to be derived by a single deletion. Among all species tested, only the related brown frog Rana dalmatina contained homologous repetitive DNA. The overall number of RgiS1a and RgiS1b repeats per R. graeca italica haploid genome was estimated to be 2.7 x 10(5). RgiS1a and RgiS1b repeats are organized in separate arrays, but repetitive units formed by various combinations of the two repeats were also observed on Southern blots. The amount of these extra repeats varies greatly among animals from the same population, representing a rare case of individual variability in the satellite DNA organization. FISH with probes specific for both satellites, or for RgiS1a only, labeled the centromeric and pericentromeric heterochromatin of all chromosomes. This indicated that RgiS1a and RgiS1b are interspersed within the same heterochromatic regions of the chromosomes.
用Asp 718I消化意大利希腊蛙(Rana graeca italica)DNA产生了281和385 bp的高度重复片段,这些片段被克隆并测序。较短的片段对应于一种卫星DNA的单位重复序列(RgiS1b)。较长的片段被发现是另一种卫星DNA(RgiS1a)的494-bp重复序列的一部分,该重复序列作为EcoRV片段被完整克隆。RgiS1b与RgiS1a有97%的同源性,似乎是由一次单碱基缺失从RgiS1a衍生而来。在所有测试的物种中,只有相关的棕蛙(Rana dalmatina)含有同源重复DNA。估计每个意大利希腊蛙单倍体基因组中RgiS1a和RgiS1b重复序列的总数为2.7×10⁵。RgiS1a和RgiS1b重复序列以单独的阵列形式组织,但在Southern杂交中也观察到由这两种重复序列的各种组合形成的重复单元。这些额外重复序列的数量在同一群体的动物中差异很大,这代表了卫星DNA组织中罕见的个体变异情况。用针对这两种卫星或仅针对RgiS1a的探针进行荧光原位杂交(FISH),标记了所有染色体的着丝粒和着丝粒周围异染色质。这表明RgiS1a和RgiS1b散布在染色体的同一异染色质区域内。