Schwarzacher Trude
Department of Biology, University of Leicester, Leicester, UK.
Methods Mol Biol. 2009;478:227-46. doi: 10.1007/978-1-59745-379-0_14.
Fluorescent chromosome analysis technologies have advanced our understanding of genome organization during the last 30 years and have enabled the investigation of DNA organization and structure as well as the evolution of chromosomes. Fluorescent chromosome staining allows even small chromosomes to be visualized, characterized by their composition and morphology, and counted. Aneuploidies and polyploidies can be established for species, breeding lines, and individuals, including changes occurring during hybridization or tissue culture and transformation protocols. Fluorescent in situ hybridization correlates molecular information of a DNA sequence with its physical location on chromosomes and genomes. It thus allows determination of the physical position of sequences and often is the only means to determine the abundance and distribution of DNA sequences that are difficult to map with any other molecular method or would require segregation analysis, in particular multicopy or repetitive DNA. Equally, it is often the best way to establish the incorporation of transgenes, their numbers, and physical organization along chromosomes. This chapter presents protocols for probe and chromosome preparation, fluorescent in situ hybridization, chromosome staining, and the analysis of results.
在过去30年里，荧光染色体分析技术增进了我们对基因组组织的理解，使人们能够研究DNA的组织和结构以及染色体的进化。荧光染色体染色能够让即使是小染色体也得以可视化，根据其组成和形态进行表征并计数。可以确定物种、育种系和个体的非整倍体和多倍体情况，包括杂交、组织培养和转化过程中发生的变化。荧光原位杂交将DNA序列的分子信息与其在染色体和基因组上的物理位置相关联。因此，它能够确定序列的物理位置，而且往往是确定那些难以用任何其他分子方法进行定位或需要进行分离分析（特别是多拷贝或重复DNA）的DNA序列的丰度和分布的唯一手段。同样，它常常是确定转基因整合情况、其数量以及沿染色体的物理组织的最佳方法。本章介绍了探针和染色体制备、荧光原位杂交、染色体染色以及结果分析的方案。
