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Visualization of the terminal structure of rice chromosomes 6 and 12 with multicolor FISH to chromosomes and extended DNA fibers
Authors:Ohmido  Nobuko  Kijima  Kyoko  Ashikawa  Ikuo  de Jong  J Hans  Fukui  Kiichi
Institution:(1) National Agricultural Research Center, Joetsu, 943-0193, Japan;(2) Laboratory of Genetics, Wageningen University, Dreijenlaan 2, 6703 HA Wageningen, The Netherlands;(3) Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, 565-0871 Osaka, Japan
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.
Keywords:extended DNA fibers (EDFs)  multicolor fluorescence in situ hybridization (McFISH)  Oryza sativa L    subtelomeric repeats  telomeres
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