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利用酵母人工染色体(YACs)对水稻第1号染色体进行物理图谱构建。

Physical mapping of rice chromosome 1 with yeast artificial chromosomes (YACs).

作者信息

Wang Z X, Idonuma A, Umehara Y, van Houten W, Ashikawa I, Minobe Y, Kurata N, Sasaki T

机构信息

Rice Genome Research Program, National Institute of Agrobiological Resources/Society for Techno-innovation of Agriculture, Forestry and Fisheries, Ibaraki, Japan.

出版信息

DNA Res. 1996 Oct 31;3(5):291-6. doi: 10.1093/dnares/3.5.291.

Abstract

We have constructed a physical map of rice chromosome 1 using yeast artificial chromosomes (YACs). A YAC library of 350 kb average insert size, covering about 6 rice haploid genome equivalents, was screened using 182 DNA markers which we had previously located on chromosome 1, by colony hybridization and polymerase chain reaction (PCR) amplification. One hundred and sixty-two DNA markers identified at least one YAC each carrying one, two or more marker sequences, for a total of 476 clones. Of these identified YACs, 284 were located in their original positions on chromosome 1. These 284 YACs defined 69 YAC contigs or islands which are estimated to cover more than 60% of the total chromosome length. The use of mapped DNA markers in constructing a physical map facilitates the integration of genetic and physical maps, as well as fine ordering of the DNA markers, especially at sites where the markers are clustered tightly on the genetic map. Our high density molecular map has been proven, by chromosome landing with YACs using mapped DNA markers, to cover more than half of the entire length of chromosome 1. The remaining 192 YACs were selected by other copies of DNA markers that mapped on chromosome 1. This description of the YAC contigs formed on chromosome 1 constitutes the second report of rice physical mapping, following that for chromosome 6.

摘要

我们利用酵母人工染色体(YAC)构建了水稻第1号染色体的物理图谱。使用平均插入片段大小为350 kb、覆盖约6个水稻单倍体基因组当量的YAC文库,通过菌落杂交和聚合酶链反应(PCR)扩增,用我们先前定位在第1号染色体上的182个DNA标记进行筛选。162个DNA标记各自鉴定出至少一个携带一个、两个或更多标记序列的YAC,总共476个克隆。在这些鉴定出的YAC中,284个位于第1号染色体上的原始位置。这284个YAC定义了69个YAC重叠群或片段,估计覆盖了染色体总长度的60%以上。在构建物理图谱时使用已定位的DNA标记有助于遗传图谱和物理图谱的整合,以及DNA标记的精细排序,特别是在遗传图谱上标记紧密聚集的位点。我们的高密度分子图谱已通过使用已定位的DNA标记以YAC进行染色体着陆的方法得到验证,覆盖了第1号染色体全长的一半以上。其余192个YAC是通过定位在第1号染色体上的DNA标记的其他拷贝筛选出来的。对第1号染色体上形成的YAC重叠群的这一描述是继第6号染色体之后水稻物理图谱构建的第二篇报道。

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