Lee H, Choi E, Seomun Y, Montgomery K, Huebner A, Lee E, Lau S, Joo C K, Kucherlapati R, Yoon S J
Research Institute of Molecular Genetics, Catholic Research Institutes of Medical Sciences, Seoul, Korea.
Genome Res. 2000 Oct;10(10):1561-7. doi: 10.1101/gr.142100.
For those searching for human disease-causing genes, information on the position of genes with respect to genetic markers is essential. The physical map composed of ESTs and genetic markers provides the positional information of these markers as well as the starting point of gene identification in the form of genomic clones containing exons. To facilitate the effort of identification of genes in the region spanning D12S1629 and D12S312, we constructed a high-resolution transcript map with PAC/BAC/cosmid clones. The strategy for the construction of such a map involved utilization of STSs for the screening of the large insert bacterial chromosome libraries and a chromosome 12-specific cosmid library by hybridization. The contig was constructed based on the STS contents of the clones. The resulting high-resolution transcript map of the region between P273P14/SP6 and D12S312 spans 4.4 cM from 66.8 to 71.2 cM of the Généthon genetic map and represents approximately 2.4 Mb. It was composed of 81 BAC, 45 PAC, and 91 cosmid clones with a minimal tiling path consisting of 16 BAC and 4 PAC clones. These clones are being used to sequence this part of chromosome 12. We determined the order of 135 STSs including 74 genes and ESTs in the map. Among these, 115 STSs were unambiguously ordered, resulting in one ordered marker per 21 kb. The order of keratin type II locus genes was determined. This map would greatly enhance the positional cloning effort of the responsible genes for those diseases that are linked to this region, including male germ cell tumor as well as palmoplantar keratoderma, Bothnian-type, and triple A syndrome. This transcript map was localized at human chromosome 12q13.
对于那些寻找人类致病基因的人来说，基因相对于遗传标记位置的信息至关重要。由EST和遗传标记组成的物理图谱提供了这些标记的位置信息，以及以包含外显子的基因组克隆形式的基因鉴定起点。为了便于在跨越D12S1629和D12S312的区域中鉴定基因，我们用PAC/BAC/黏粒克隆构建了一个高分辨率转录图谱。构建这样一个图谱的策略包括利用STS通过杂交筛选大插入片段细菌染色体文库和12号染色体特异性黏粒文库。重叠群是根据克隆的STS含量构建的。所得的P273P14/SP6和D12S312之间区域的高分辨率转录图谱在Généthon遗传图谱的66.8至71.2 cM范围内跨越4.4 cM，约为2.4 Mb。它由81个BAC、45个PAC和91个黏粒克隆组成，最小重叠路径由16个BAC和4个PAC克隆组成。这些克隆正用于对12号染色体的这一部分进行测序。我们确定了该图谱中135个STS的顺序，其中包括74个基因和EST。其中，115个STS的顺序明确，平均每21 kb有一个有序标记。确定了角蛋白II型基因座基因的顺序。该图谱将大大加强对与该区域相关疾病（包括男性生殖细胞肿瘤以及波的尼亚型掌跖角化病和三A综合征）致病基因的定位克隆工作。该转录图谱定位于人类12号染色体q13区。
