对来自人类染色体着丝粒区域大小超过5.7兆碱基的DNA限制片段的分析。

Analysis of DNA restriction fragments greater than 5.7 Mb in size from the centromeric region of human chromosomes.

作者信息

Arn P H, Li X, Smith C, Hsu M, Schwartz D C, Jabs E W

机构信息

Department of Pediatrics, Johns Hopkins University, Baltimore, Maryland 21205.

出版信息

Mamm Genome. 1991;1(4):249-54. doi: 10.1007/BF00352332.

DOI:10.1007/BF00352332

PMID:1665375

Abstract

Pulsed electrophoresis was used to study the organization of the human centromeric region. Genomic DNA was digested with rare-cutting enzymes. DNA fragments from 0.2 to greater than 5.7 Mb were separated by electrophoresis and hybridized with alphoid and simple DNA repeats. Rare-cutting enzymes (Mlu I, Nar I, Not I, Nru I, Sal I, Sfi I, Sst II) demonstrated fewer restriction sites at centromeric regions than elsewhere in the genome. The enzyme Not I had the fewest restriction sites at centromeric regions. As much as 70% of these sequences from the centromeric region are present in Not I DNA fragments greater than 5.7 and estimated to be as large as 10 Mb in size. Other repetitive sequences such as short interspersed repeated segments (SINEs), long interspersed repeated segments (LINEs), ribosomal DNA, and mini-satellite DNA that are not enriched at the centromeric region, are not enriched in Not I fragments of greater than 5.7 Mb in size.

摘要

脉冲电泳用于研究人类着丝粒区域的组织。基因组DNA用稀有切割酶消化。0.2至大于5.7兆碱基的DNA片段通过电泳分离，并用α卫星DNA和简单DNA重复序列杂交。稀有切割酶（Mlu I、Nar I、Not I、Nru I、Sal I、Sfi I、Sst II）在着丝粒区域的限制酶切位点比基因组其他地方少。Not I酶在着丝粒区域的限制酶切位点最少。来自着丝粒区域的这些序列中多达70%存在于大于5.7兆碱基的Not I DNA片段中，估计大小达10兆碱基。其他重复序列，如短散在重复片段（SINEs）、长散在重复片段（LINEs）、核糖体DNA和小卫星DNA，在着丝粒区域不富集，在大小大于5.7兆碱基的Not I片段中也不富集。