Schmidt R, Love K, West J, Lenehan Z, Dean C
Department of Molecular Genetics, John Innes Centre, Colney, Norwich, UK.
Plant J. 1997 Mar;11(3):563-72. doi: 10.1046/j.1365-313x.1997.11030563.x.
In order to generate a physical map of Arabidopsis thaliana chromosome 5, 142 molecular markers mapping chromosome 5 have been used in colony hybridization experiments with four Arabidopsis, ecotype Columbia, yeast artificial chromosome (YAC) libraries. This resulted in 634 YAC clones being anchored on chromosome 5. Southern blot analysis confirmed their positioning and provided data, which along with knowledge of the sizes of all the YAC clones, enabled the clones to be arranged into 31 contigs. Genetic mapping of markers located within 29 of these contigs on the Landsberg erecta/Columbia recombinant inbred lines allowed positioning of the contigs along the chromosome. A high proportion of the YAC clones were found to contain chimaeric inserts. The availability of this YAC contig map will accelerate chromosome-walking experiments, provide substrates for large-scale genomic sequencing projects and facilitate the mapping of new probes to this chromosome.
为了构建拟南芥第5号染色体的物理图谱,在对4个拟南芥生态型哥伦比亚酵母人工染色体(YAC)文库进行菌落杂交实验时,使用了142个定位在第5号染色体上的分子标记。这使得634个YAC克隆被锚定在第5号染色体上。Southern杂交分析证实了它们的定位,并提供了相关数据,结合所有YAC克隆的大小信息,能够将这些克隆排列成31个重叠群。对这些重叠群中29个所含标记在Landsberg erecta/哥伦比亚重组自交系上进行遗传作图,从而将重叠群定位到染色体上。发现高比例的YAC克隆含有嵌合插入片段。这个YAC重叠群图谱的可用性将加速染色体步移实验,为大规模基因组测序项目提供底物,并便于将新探针定位到这条染色体上。