Brunkow M E, Nagle D L, Bernstein A, Bucan M
Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
Genomics. 1995 Jan 20;25(2):421-32. doi: 10.1016/0888-7543(95)80042-k.
We constructed a yeast artificial chromosome (YAC) contig spanning the genes encoding Kit (Kit), the platelet-derived growth factor alpha receptor (Pdgfra), and fetal liver kinase 1 (Flk1), three members of a receptor tyrosine kinase gene family located in the central portion of mouse chromosome 5. The orientation of YAC clones and the extent of their overlap was determined by "probe content mapping," that is, hybridization analysis of YAC clones using the available gene probes and YAC end sequences. For four YAC clones, which constitute a minimal set spanning 1.8 Mb, a detailed restriction map was constructed. This map, in conjunction with the previously published long-range restriction map, indicates the order, the physical distances, and the relative transcriptional orientations of the Pdgfra, Kit, and Flk1 genes. The YAC clones and corresponding YAC end probes presented here provide an important resource for the molecular analysis of a cluster of developmental mutations, namely dominant white spotting (W), patch (Ph), recessive spotting (rs), and rump-white (Rw), associated with this chromosomal region.
我们构建了一个酵母人工染色体(YAC)重叠群,该重叠群跨越了编码Kit(Kit)、血小板衍生生长因子α受体(Pdgfra)和胎儿肝激酶1(Flk1)的基因,这三个基因是位于小鼠5号染色体中部的受体酪氨酸激酶基因家族的成员。YAC克隆的方向及其重叠程度是通过“探针含量作图”确定的,即使用可用的基因探针和YAC末端序列对YAC克隆进行杂交分析。对于构成一个跨越1.8 Mb的最小集合的四个YAC克隆,构建了详细的限制性图谱。该图谱与先前发表的长程限制性图谱一起,表明了Pdgfra、Kit和Flk1基因的顺序、物理距离和相对转录方向。本文提供的YAC克隆和相应的YAC末端探针为分子分析与该染色体区域相关的一组发育突变,即显性白斑(W)、斑块(Ph)、隐性斑点(rs)和臀部白色(Rw),提供了重要资源。