Foster J W, Schafer A J, Critcher R, Spillett D J, Feakes R W, Walter M A, Dominguez-Steglich M, Guioli S, Brook J D, Goodfellow P N
Department of Genetics, University of Cambridge, United Kingdom.
Genomics. 1996 Apr 15;33(2):185-92. doi: 10.1006/geno.1996.0182.
We have constructed a whole genome radiation hybrid (WG-RH) map across a region of human chromosome 17q, from growth hormone (GH) to thymidine kinase (TK). A panel of 128 WG-RH hybrid cell lines generated by X-irradiation and fusion has been tested for the retention of 39 sequence-tagged site (STS) markers by the polymerase chain reaction. This genome mapping technique has allowed the integration of existing VNTR and microsatellite markers with additional new markers and existing STS markers previously mapped to this region by other means. The WG-RH map includes eight expressed sequence tag (EST) and three anonymous markers developed for this study, together with 23 anonymous microsatellites and five existing ESTs. Analysis of these data resulted in a high-density comprehensive map across this region of the genome. A subset of these markers has been used to produce a framework map consisting of 20 loci ordered with odds greater than 1000:1. The markers are of sufficient density to build a YAC contig across this region based on marker content. We have developed sequence tags for both ends of a 2.1-Mb YAC and mapped these using the WG-RH panel, allowing a direct comparison of cRay6000 to physical distance.
我们构建了一张跨越人类17号染色体q区从生长激素(GH)到胸苷激酶(TK)区域的全基因组辐射杂种(WG-RH)图谱。通过X射线照射和细胞融合产生的一组128个WG-RH杂种细胞系,已通过聚合酶链反应检测了39个序列标签位点(STS)标记的保留情况。这种基因组作图技术使得现有的VNTR和微卫星标记与新的附加标记以及先前通过其他方法定位到该区域的现有STS标记得以整合。WG-RH图谱包括为本研究开发的8个表达序列标签(EST)和3个匿名标记,以及23个匿名微卫星和5个现有的EST。对这些数据的分析产生了一张跨越该基因组区域的高密度综合图谱。这些标记中的一个子集已被用于构建一个框架图谱,该图谱由20个排序概率大于1000:1的位点组成。这些标记的密度足以基于标记内容构建跨越该区域的酵母人工染色体(YAC)重叠群。我们为一个2.1兆碱基对的YAC的两端开发了序列标签,并使用WG-RH细胞系对其进行定位,从而能够直接比较cRay6000与物理距离。