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高粱着丝粒相关重复元件 CEN38 的分布与序列分析(禾本科)。

Distribution and sequence analysis of the centromere-associated repetitive element CEN38 of Sorghum bicolor (Poaceae).

机构信息

Investigen Inc., 851 West Midway Ave., Alameda, California 94501 USA;

出版信息

Am J Bot. 2000 Dec;87(12):1757-64.

Abstract

Fluorescence in situ hybridization (FISH) of a large-insert genomic clone, BAC 22B2, previously suggested that Sorghum bicolor (2n = 20) has the tetraploid architecture A(b)A(b)B(b)B(b). Here, we report on BAC 22B2 subclone pCEN38 (1047-bp insert) as related to sorghum and sugarcane. Mitotic FISH of six different subclones of BAC 22B2 showed that pCEN38 produced the strongest specificity to the A(b) subgenome and signal occurred primarily near centromeres. Southern blots of pCEN38 to 21 crop plants revealed a narrow taxonomic distribution. Meiotic metaphase I FISH positioned pCEN38 sequences near active centromeres. Pachytene FISH revealed that the distributions are trimodal in several B(b) and possibly all sorghum chromosomes. DNA sequencing revealed that the pCEN38 fragment contains three tandemly repeated dimers (<280 bp) of the same sequence family found in sorghum clone pSau3A10, and that each dimer consists of two divergent monomers (<140 bp). Sequence comparisons revealed homology between the pCEN38 monomers and the SCEN 140 bp tandem repeat family of sugarcane. FISH of pCEN38 yielded signal in centromere regions of most but not all sugarcane chromosomes. Results suggest that sugarcane and sorghum share at least one ancestor harboring elements similar to pCEN38 and SCEN and that each species had an ancestor in which the repetitive element was weakly present or lacking.

摘要

荧光原位杂交(FISH)分析一个大插入基因组克隆 BAC 22B2,先前表明高粱(2n = 20)具有四倍体结构 A(b)A(b)B(b)B(b)。在这里,我们报告了 BAC 22B2 亚克隆 pCEN38(1047-bp 插入)与高粱和甘蔗有关。六种不同 BAC 22B2 亚克隆的有丝分裂 FISH 表明,pCEN38 对 A(b)亚基因组具有最强的特异性,信号主要发生在着丝粒附近。pCEN38 对 21 种作物植物的 Southern blot 显示出狭窄的分类分布。减数分裂中期 I FISH 将 pCEN38 序列定位在活性着丝粒附近。粗线期 FISH 显示,在几个 B(b)和可能所有高粱染色体中,分布呈三峰模式。DNA 测序表明,pCEN38 片段包含三个串联重复的二聚体(<280 bp),属于在高粱克隆 pSau3A10 中发现的相同序列家族,并且每个二聚体由两个发散的单体(<140 bp)组成。序列比较表明,pCEN38 单体与甘蔗的 SCEN 140 bp 串联重复家族之间存在同源性。pCEN38 的 FISH 在大多数但不是所有甘蔗染色体的着丝粒区域产生信号。结果表明,甘蔗和高粱至少共享一个祖先,其携带与 pCEN38 和 SCEN 相似的元件,并且每个物种的祖先都存在重复元件存在或缺乏的情况。

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