Practicable approaches to facilitate rapid and accurate molecular cytogenetic mapping in birds and mammals
Practicable approaches to facilitate rapid and accurate molecular cytogenetic mapping in birds and mammals
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DOI：
10.1159/000103163
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发表时间：
2007-01-01
影响因子：
1.7
通讯作者：
Griffin, D. K.
中科院分区：
文献类型：
--
作者：
Morris, W. B.;Stephenson, J. E.;Griffin, D. K.
FISH的分子细胞遗传学定位是大多数基因组计划的共同特点，因为它提供了基因组的全球低分辨率概述，并促进了比较基因组学。细胞遗传学作图的一个基本前提是能够准确地识别克隆（如BAC）所在的染色体。由于对人类核型的了解是司空见惯的，因此这通常不会成为绘制人类染色体图谱的障碍。然而，对其他物种来说，准确的分配可能是有问题的，要么是因为像鸟类那样，核型太复杂，无法用标准方法分析，要么是因为缺乏熟练进行核型分析的个体。以鸡为模型，我们开发了一种可重复的精确细胞遗传学定位方法，包括：单色FISH，测量信号携带染色体与8号染色体的大小之比，并通过一系列小的双色实验最终分配。使用来自Ensembl浏览器的碱基对估计信息建立尺寸比的参考值。通过这种方法，细胞遗传学定位到高度复杂的核型可以实现在少数简单的步骤。我们还开发并测试了一个核型教程程序，该程序改编自本杂志先前报道的一个程序。也就是说，我们已经用猪作为一个具有相对易处理核型的模式物种的例子，并证明科学家和学生，即使只使用我们的教程一个小时，也可以很容易地识别猪的染色体，从而使用FISH进行适当的分配。对于科学问题的解决，简单可行的方法往往比复杂的方法（例如m-FISH）提供更好的解决方案。这就是本文描述的方法的情况。版权所有(c) 2007 S. Karger AG，巴塞尔
Molecular cytogenetic mapping by FISH is a common feature of most genome projects as it provides a global, low-resolution overview of the genome and facilitates comparative genomics. An essential prerequisite for cytogenetic mapping is the ability to identify accurately the chromosome on which the clone ( e. g. BAC) resides. This is not usually a barrier to human mapping as knowledge of the human karyotype is commonplace. For other species however accurate assignment can be problematic either because, as in birds, the karyotype is too complex to analyze by standard means or because of the paucity of individuals skilled to perform the karyotyping. Using chicken as a model we have developed a reproducible approach for accurate cytogenetic mapping that involves: a single colour FISH, measurement of the ratio of the size of the signal bearing chromosome to that of chromosome 8, and final assignment through a small series of dual colour experiments. Reference values for size ratios were established using base pair estimate information from the Ensembl browser. By this method cytogenetic mapping to highly complex karyotypes can be achieved in a small number of simple steps. We have also developed and tested a karyotyping tutorial programme adapted from one previously reported in this journal. That is, we have used pig as an example of a model species with a relatively tractable karyotype and demonstrated that scientists and students, even after only one hour using our tutorial, can readily identify pig chromosomes and thus make appropriate assignments using FISH. Simple, practicable means often provide preferable solutions than complex alternatives ( e. g. m-FISH) to the solution of scientific problems. Such is the case for the approaches described here. Copyright (c) 2007 S. Karger AG, Basel