Brinkmeyer-Langford C, Raudsepp T, Gustafson-Seabury A, Chowdhary B P
Department of Veterinary Integrative Biomedical Sciences, College of Veterinary Medicine, Texas A&M University, College Station, TX, USA.
Cytogenet Genome Res. 2008;120(1-2):164-72. doi: 10.1159/000118758. Epub 2008 Apr 30.
A total of 207 BAC clones containing 155 loci were isolated and arranged into a map of linearly ordered overlapping clones over the proximal part of horse chromosome 21 (ECA21), which corresponds to the proximal half of the short arm of human chromosome 19 (HSA19p) and part of HSA5. The clones form two contigs - each corresponding to the respective human chromosomes - that are estimated to be separated by a gap of approximately 200 kb. Of the 155 markers present in the two contigs, 141 (33 genes and 108 STS) were generated and mapped in this study. The BACs provide a 4-5x coverage of the region and span an estimated length of approximately 3.3 Mb. The region presently contains one mapped marker per 22 kb on average, which represents a major improvement over the previous resolution of one marker per 380 kb obtained through the generation of a dense RH map for this segment. Dual color fluorescence in situ hybridization on metaphase and interphase chromosomes verified the relative order of some of the BACs and helped to orient them accurately in the contigs. Despite having similar gene order and content, the equine region covered by the contigs appears to be distinctly smaller than the corresponding region in human (3.3 Mb vs. 5.5-6 Mb) because the latter harbors a host of repetitive elements and gene families unique to humans/primates. Considering limited representation of the region in the latest version of the horse whole genome sequence EquCab2, the dense map developed in this study will prove useful for the assembly and annotation of the sequence data on ECA21 and will be instrumental in rapid search and isolation of candidate genes for traits mapped to this region.
共分离出207个包含155个位点的BAC克隆,并将它们排列成马21号染色体(ECA21)近端部分的线性有序重叠克隆图谱,该区域对应于人类19号染色体短臂的近端一半(HSA19p)和5号染色体的一部分。这些克隆形成了两个重叠群,每个重叠群分别对应于各自的人类染色体,据估计它们之间相隔约200 kb的间隙。在这两个重叠群中存在的155个标记中,有141个(33个基因和108个STS)是在本研究中产生并定位的。这些BAC克隆对该区域的覆盖度为4-5倍,跨度估计约为3.3 Mb。目前该区域平均每22 kb有一个定位标记,这比之前通过为该片段生成密集的RH图谱获得的每380 kb一个标记的分辨率有了显著提高。对中期和间期染色体进行的双色荧光原位杂交验证了一些BAC克隆的相对顺序,并有助于在重叠群中准确地对它们进行定位。尽管重叠群覆盖的马区域具有相似的基因顺序和内容,但由于人类/灵长类特有的大量重复元件和基因家族,该区域似乎明显小于人类相应区域(3.3 Mb对5.5-6 Mb)。考虑到最新版马全基因组序列EquCab2中该区域的代表性有限,本研究中开发的密集图谱将被证明对ECA21序列数据的组装和注释有用,并将有助于快速搜索和分离定位到该区域的性状的候选基因。