Ichikawa K, Shimamoto A, Imamura O, Tokutake Y, Yamabe Y, Kitao S, Suzuki N, Sugawara K, Matsumoto T, Thomas W, Drayna D, Goto M, Sugimoto M, Sugawara M, Furuichi Y
AGENE Research Institute, Kanagawa, Japan.
DNA Res. 1998 Apr 30;5(2):103-13. doi: 10.1093/dnares/5.2.103.
Detailed physical maps of the human genome are important resources for identification and isolation of genes responsible for diseases and for the study of their structure and function. We constructed a 2.0-Mb high-resolution physical map within the human chromosome 8p12-p21 region extending from marker D8S131 to D8S283. The map comprises a series of contigs mostly P1/PAC clones, which span the loci of potential tumor suppressor genes and the Werner's syndrome gene. Each P1/PAC DNA was defined by its size, restriction sites, terminal sequences, intermarker distances and location relative to major genes and markers. The genes on these P1/PAC DNAs were analyzed by an exon amplification method to determine their locations. The genes newly found by the exon amplification method together with other known genes, including those of glutathion reductase, a general transcription factor, protein phosphatase 2A beta subunit and Werner's syndrome, were precisely mapped within the contigs. These P1/PAC DNAs are useful reagents for the generation of new microsatellite markers to narrow the candidate region of the tumor suppressor gene(s) and/or genes responsible for other diseases, which are believed to exist in this region by linkage analysis.
人类基因组的详细物理图谱是鉴定和分离致病基因以及研究其结构与功能的重要资源。我们在人类染色体8p12 - p21区域构建了一个2.0兆碱基对的高分辨率物理图谱,该区域从标记D8S131延伸至D8S283。该图谱由一系列重叠群组成,大多是P1/PAC克隆,它们跨越了潜在肿瘤抑制基因和沃纳综合征基因的位点。每个P1/PAC DNA通过其大小、限制性酶切位点、末端序列、标记间距离以及相对于主要基因和标记的位置来定义。通过外显子扩增方法分析这些P1/PAC DNA上的基因,以确定它们的位置。通过外显子扩增方法新发现的基因以及其他已知基因,包括谷胱甘肽还原酶、一种通用转录因子、蛋白磷酸酶2Aβ亚基和沃纳综合征的基因,都被精确地定位在重叠群内。这些P1/PAC DNA是用于生成新微卫星标记的有用试剂,以缩小肿瘤抑制基因和/或其他致病基因的候选区域,通过连锁分析认为这些基因存在于该区域。