Umehara Y, Tanoue H, Kurata N, Ashikawa I, Minobe Y, Sasaki T
Rice Genome Research Program, National Institute of Agrobiological Resources/Society for Techno-innovation of Agriculture, Forestry and Fisheries, Ibaraki, Japan.
Genome Res. 1996 Oct;6(10):935-42. doi: 10.1101/gr.6.10.935.
Yeast artificial chromosome (YAC) clones carrying DNA marker sequences located on the rice genetic map of chromosome 6 were ordered for physical mapping. A total of 122 restriction fragment length polymorphism markers, 16 sequence-tagged site markers, and five random amplified polymorphic DNA markers located, on average, at 0.9-cM intervals, were used for YAC clone screening by colony/Southern hybridization and PCR screening, respectively. A total of 216 individual YACs were selected from our YAC library of 7000 clones covering six genome equivalents. Each DNA marker could select, on average, 4.8 YAC clones, with 11 clones being the maximum. The YACs localized to the corresponding linkage map positions form 43 contigs and encompass about 60% of rice chromosome 6. This is the first step in constructing a physical map covering the whole rice genome by chromosome landing with YAC clones. These YACs and data will be used soon to isolate phenotypical trait genes by map-based cloning.
对携带位于水稻第6号染色体遗传图谱上的DNA标记序列的酵母人工染色体(YAC)克隆进行排序,以构建物理图谱。分别使用平均间隔为0.9厘摩定位的总共122个限制性片段长度多态性标记、16个序列标签位点标记和5个随机扩增多态性DNA标记,通过菌落/ Southern杂交和PCR筛选来筛选YAC克隆。从我们包含六个基因组当量的7000个克隆的YAC文库中总共挑选出216个单个YAC。每个DNA标记平均可以挑选出4.8个YAC克隆,最多为11个。定位到相应连锁图谱位置的YAC形成了43个重叠群,覆盖了水稻第6号染色体约60%的区域。这是以YAC克隆通过染色体登陆构建覆盖整个水稻基因组的物理图谱的第一步。这些YAC和数据很快将用于通过图位克隆分离表型性状基因。