Nanda I, Zischler H, Epplen C, Guttenbach M, Schmid M
Department of Human Genetics, University of Würzburg, Germany.
Electrophoresis. 1991 Feb-Mar;12(2-3):193-203. doi: 10.1002/elps.1150120216.
Stretches of short, simple DNA sequences are widespread in all eukaryote genomes studied so far. Simple sequences are thought to undergo frequent expansion and deletion due to intrinsic genomic mechanisms. Some of the simple sequences were used successfully to detect hypervariable loci in various genomes. Hybridization experiments using synthetic probes not only revealed the informative simple repeats suitable for DNA fingerprinting in a particular species, but also reflected the wide range of distribution of the simple sequences among eukaryotes. The organization of these simple repetitive sequences at the chromosomal loci was investigated using in situ hybridization with chemically synthesized, pure oligonucleotide probes. Both biotin- and digoxigenin-attached probes detected specific chromosomal sites that are enriched in the respective simple-repeat blocks. Depending on the organism and probe used, accumulation of simple DNA sequences at individual or multiple sites on the chromosomes of different vertebrates could be demonstrated. The simple repetitive DNA sequences are located in different chromosomal regions (e.g., heterochromatin on the sex chromosomes, nucleolus organizer regions, and R-band sites), which are constrained considerably during evolution.
到目前为止,短的、简单的DNA序列片段在所有已研究的真核生物基因组中广泛存在。由于内在的基因组机制,简单序列被认为会频繁发生扩增和缺失。一些简单序列已成功用于检测各种基因组中的高变位点。使用合成探针进行的杂交实验不仅揭示了适合特定物种DNA指纹分析的信息丰富的简单重复序列,还反映了简单序列在真核生物中的广泛分布。利用化学合成的纯寡核苷酸探针进行原位杂交,研究了这些简单重复序列在染色体位点的组织情况。生物素标记和地高辛配基标记的探针均检测到特定的染色体位点,这些位点富含各自的简单重复序列块。根据所使用的生物和探针的不同,可以证明不同脊椎动物染色体上的单个或多个位点存在简单DNA序列的积累。简单重复DNA序列位于不同的染色体区域(例如,性染色体上的异染色质、核仁组织区和R带位点),这些区域在进化过程中受到很大限制。