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利用染色体和伸展DNA纤维的多色荧光原位杂交技术对水稻6号和12号染色体的末端结构进行可视化分析。

Visualization of the terminal structure of rice chromosomes 6 and 12 with multicolor FISH to chromosomes and extended DNA fibers.

作者信息

Ohmido N, Kijima K, Ashikawa I, de Jong J H, Fukui K

机构信息

National Agricultural Research Center, Joetsu, Japan.

出版信息

Plant Mol Biol. 2001 Oct;47(3):413-21. doi: 10.1023/a:1011632111845.

Abstract

High-resolution fluorescence in situ hybridization (FISH) on interphase and pachytene nuclei, and extended DNA fibers enabled microscopic distinction of DNA sequences less than a few thousands of base pairs apart. We applied this technique to reveal the molecular organization of telomere ends in japonica rice (Oryza sativa ssp. japonica), which consist of the Arabidopsis type TTTAGGG heptameric repeats and the rice specific subtelomeric tandem repeat sequence A (TrsA). Southern hybridizations of DNA digested with Bal31 and EcoRI, and FISH on chromosomes and extended DNA fibers demonstrated that (1) all chromosome ends possess the telomere tandem repeat measuring 3-4 kb; (2) the subtelomeric TrsA occurs only at the ends of the long arms of chromosomes 6 and 12, and measure 6 and 10 kb, which corresponds to 231 and 682 copies for these sites, respectively; (3) the telomere and TrsA repeats are separated by at most a few thousands of intervening nucleotide sequences. The molecular organization for a general telomere organization in plant chromosomes is discussed.

摘要

对间期和粗线期细胞核进行高分辨率荧光原位杂交(FISH),以及对伸展的DNA纤维进行杂交,能够在显微镜下区分相隔不到几千个碱基对的DNA序列。我们应用这项技术来揭示粳稻(Oryza sativa ssp. japonica)端粒末端的分子组织,其由拟南芥型TTTAGGG七聚体重复序列和水稻特有的亚端粒串联重复序列A(TrsA)组成。用Bal31和EcoRI消化的DNA进行Southern杂交,以及在染色体和伸展的DNA纤维上进行FISH,结果表明:(1)所有染色体末端都具有长度为3 - 4 kb的端粒串联重复序列;(2)亚端粒TrsA仅出现在6号和12号染色体长臂的末端,长度分别为6 kb和10 kb,这些位点分别对应231个和682个拷贝;(3)端粒和TrsA重复序列之间最多相隔几千个间隔核苷酸序列。本文还讨论了植物染色体中一般端粒组织的分子组织情况。

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