Yang F, Milne B S, Schelling C, Dolf G, Schläpfer J, Switonski M, Ladon D, Pienkowska A, Bosma A A, Sargan D R, Ferguson-Smith M A
Centre for Veterinary Science, Department of Clinical Veterinary Medicine, University of Cambridge, UK.
Chromosome Res. 2000;8(2):93-100. doi: 10.1023/a:1009251917072.
We have developed a novel method for identifying dog chromosomes and unambiguously mapping specific clones onto canine chromosomes. This method uses a previously established red fox/dog comparative chromosome map to guide the FISH mapping of cloned canine DNA. Mixing metaphase preparations of the red fox and dog enabled a single hybridization to be performed on both species. We used this approach to map the chromosomal locations of twenty-six canine cosmids. Each cosmid contains highly polymorphic microsatellite markers currently used by the DogMap project to compile the canine linkage map. All but two cosmids were successfully assigned to subchromosomal regions on red fox and dog chromosomes. For eight cosmids previously mapped on dog chromosomes, we confirmed and refined the canine chromosomal assignments of seven cosmids and corrected an erroneous assignment regarding cosmid CanBern1. These results demonstrate that the red fox and dog comparative chromosome map can greatly improve the accuracy and efficiency of chromosomal assignments of canine genetic markers by FISH.
我们开发了一种鉴定犬类染色体并将特定克隆明确地定位到犬类染色体上的新方法。该方法利用先前建立的赤狐/犬类比较染色体图谱来指导克隆犬类DNA的荧光原位杂交(FISH)定位。将赤狐和犬类的中期染色体标本混合,使得能够在两个物种上同时进行一次杂交。我们采用这种方法来定位26个犬类黏粒的染色体位置。每个黏粒都包含目前犬类图谱项目用于编制犬类连锁图谱的高度多态性微卫星标记。除了两个黏粒外,所有黏粒都成功地定位到了赤狐和犬类染色体的亚染色体区域。对于先前已定位在犬类染色体上的8个黏粒,我们确认并细化了其中7个黏粒的犬类染色体定位,并纠正了关于黏粒CanBern1的错误定位。这些结果表明,赤狐和犬类比较染色体图谱可以通过FISH大大提高犬类遗传标记染色体定位的准确性和效率。