用染色体特异性BAC克隆绘制拟南芥染色体

Q1 Agricultural and Biological Sciences Current protocols in plant biology Pub Date : 2016-06-10 DOI:10.1002/cppb.20022

Terezie Mandáková, Martin A. Lysak

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引用次数: 39

摘要

染色体彩绘(CP)是指对染色体特异性DNA探针进行荧光原位杂交(FISH)，以识别大的染色体区域、染色体臂和整个染色体。对于植物的CP和CCP(比较染色体绘制)，通常使用来自原物种或近缘物种的染色体特异性细菌人工染色体(BAC)群作为绘制探针。CP能够可视化和追踪特定的染色体区域和/或染色体在所有有丝分裂和减数分裂阶段以及相应的间期染色体区域。CCP能够鉴定大尺度同源染色体区域和两个或多个物种之间共享的染色体。本文描述了在拟南芥(Arabidopsis thaliana)和其他十字花科植物分类群中使用拟南芥染色体特异性BAC contigs进行CP和CCP的逐步方案。©2016 by John Wiley &儿子,Inc。

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Painting of Arabidopsis Chromosomes with Chromosome-Specific BAC Clones

Chromosome painting (CP) refers to fluorescence in situ hybridization (FISH) of chromosome-specific DNA probes to identify large chromosome regions, chromosome arms, and whole chromosomes. For CP and CCP (comparative chromosome painting) in plants, most often, contigs of chromosome-specific bacterial artificial chromosomes (BAC) from the species of origin or a related species are used as painting probes. CP enables visualization and tracing of particular chromosome regions and/or chromosomes throughout all mitotic and meiotic stages as well as the corresponding interphase chromosome territories. CCP enables identification of large-scale homeologous chromosome regions and chromosomes shared among two or more species. Here, a step-by-step protocol for carrying out CP in Arabidopsis thaliana (Arabidopsis) and CCP in other crucifer taxa based on the use of Arabidopsis chromosome-specific BAC contigs is described. © 2016 by John Wiley & Sons, Inc.

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Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.