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[水稻属不同基因组着丝粒BAC克隆的分离与鉴定]

[Isolation and characterization of the centromeric BAC clones from different genomes in genus Oryza].

作者信息

Yi Chuan-Deng, Gong Zhi-Yun, Liang Guo-Hua, Wang Fei-Hua, Tang Shu-Zhu, Gu Ming-Hong

机构信息

Key Lab of Plant Functional Genomics, Ministry of Education, Yangzhou University, Jiangsu Province Key Lab of Crop Genetics and Physiology, Yangzhou 225009, China.

出版信息

Yi Chuan. 2007 Jul;29(7):851-8. doi: 10.1360/yc-007-0851.

Abstract

Centromeres play an important role in ensuring the correct segregation and transmission of chromosome during mitosis and meiosis in eukaryotes. In this research, we constructed five BAC libraries for diploid wild rice with different genomes. Together with the technique of colony blot hybridization and fluorescence in situ hybridization (FISH), centromere-related BAC clones were screened and characterized from different genomes. Meanwhile, co-hybridization was detected between these clones and the five genomes. The results from this study demonstrated that: (1) there were centromere-specific satellite repeat in Oryza officinalis (CC genome) and O. brachyantha (FF genome), respectively, and centromere-specific CRR-related sequence was found in O. brachyantha; (2) homology sequences of CentO and CRR of O. sativa (AA genome) were detected on all centromeres of O. glaberrima (AA genome), O. punctata (BB genome) and O. australiensis (EE genome); And (3) the two somatic chromosomes of O. officinalis comprised of homology sequences of CentO satellites as revealed FISH analysis probed with RCS2. Homology sequences of CRR of O. sativa were also detected on all centromeres of O. officinalis. The results provided a foundation toward cloning the centromeric sequences from different genomes of genus Oryza, studying centromere organization and evolution of different genome, analyzing the relationship between centromeric structure and function among different genome.

摘要

着丝粒在真核生物有丝分裂和减数分裂过程中确保染色体的正确分离和传递方面发挥着重要作用。在本研究中,我们构建了五个不同基因组的二倍体野生稻BAC文库。结合菌落杂交和荧光原位杂交(FISH)技术,从不同基因组中筛选并鉴定了与着丝粒相关的BAC克隆。同时,检测了这些克隆与五个基因组之间的共杂交情况。本研究结果表明:(1)药用野生稻(CC基因组)和短药野生稻(FF基因组)分别存在着丝粒特异性卫星重复序列,并且在短药野生稻中发现了着丝粒特异性CRR相关序列;(2)在光稃稻(AA基因组)、斑点野生稻(BB基因组)和澳洲野生稻(EE基因组)的所有着丝粒上均检测到了水稻(AA基因组)CentO和CRR的同源序列;(3)用RCS2进行FISH分析显示,药用野生稻的两条体细胞染色体由CentO卫星的同源序列组成。在药用野生稻的所有着丝粒上也检测到了水稻CRR的同源序列。这些结果为从稻属不同基因组克隆着丝粒序列、研究不同基因组着丝粒的组织和进化、分析不同基因组着丝粒结构与功能之间的关系奠定了基础。

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