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番茄主要重复序列的荧光原位杂交定位及分子组织分析

FISH mapping and molecular organization of the major repetitive sequences of tomato.

作者信息

Chang Song-Bin, Yang Tae-Jin, Datema Erwin, van Vugt Joke, Vosman Ben, Kuipers Anja, Meznikova Marie, Szinay Dóra, Lankhorst René Klein, Jacobsen Evert, de Jong Hans

机构信息

Wageningen University, Laboratory of Genetics, 6703 BD, Wageningen, The Netherlands.

出版信息

Chromosome Res. 2008;16(7):919-33. doi: 10.1007/s10577-008-1249-z. Epub 2008 Aug 13.

Abstract

This paper presents a bird's-eye view of the major repeats and chromatin types of tomato. Using fluorescence in-situ hybridization (FISH) with Cot-1, Cot-10 and Cot-100 DNA as probes we mapped repetitive sequences of different complexity on pachytene complements. Cot-100 was found to cover all heterochromatin regions, and could be used to identify repeat-rich clones in BAC filter hybridization. Next we established the chromosomal locations of the tandem and dispersed repeats with respect to euchromatin, nucleolar organizer regions (NORs), heterochromatin, and centromeres. The tomato genomic repeats TGRII and TGRIII appeared to be major components of the pericentromeres, whereas the newly discovered TGRIV repeat was found mainly in the structural centromeres. The highly methylated NOR of chromosome 2 is rich in GACA, a microsatellite that also forms part of the pericentromeres, together with GA, GATA and Ty1-copia. Based on the morphology of pachytene chromosomes and the distribution of repeats studied so far, we now propose six different chromatin classes for tomato: (1) euchromatin, (2) chromomeres, (3) distal heterochromatin and interstitial heterochromatic knobs, (4) pericentromere heterochromatin, (5) functional centromere heterochromatin and (6) nucleolar organizer region.

摘要

本文概述了番茄的主要重复序列和染色质类型。我们使用以Cot-1、Cot-10和Cot-100 DNA为探针的荧光原位杂交(FISH)技术,在粗线期染色体上绘制了不同复杂程度的重复序列图谱。发现Cot-100覆盖了所有异染色质区域,可用于在BAC滤膜杂交中鉴定富含重复序列的克隆。接下来,我们确定了串联重复序列和分散重复序列相对于常染色质、核仁组织区(NORs)、异染色质和着丝粒的染色体位置。番茄基因组重复序列TGRII和TGRIII似乎是着丝粒周围区域的主要成分,而新发现的TGRIV重复序列主要存在于结构着丝粒中。2号染色体高度甲基化的NOR富含GACA,这是一种微卫星序列,它与GAGATA和Ty1- copia一起也是着丝粒周围区域的一部分。根据粗线期染色体的形态以及目前所研究的重复序列的分布,我们现在为番茄提出六种不同的染色质类别:(1)常染色质,(2)染色粒,(3)远端异染色质和间期异染色质结,(4)着丝粒周围异染色质,(5)功能性着丝粒异染色质,(6)核仁组织区。

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