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水稻第5号染色体上酵母人工染色体连续克隆系的构建。

Construction of YAC contigs on rice chromosome 5.

作者信息

Saji S, Umehara Y, Kurata N, Ashikawa I, 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):297-302. doi: 10.1093/dnares/3.5.297.

Abstract

A physical map of rice chromosome 5 was constructed with yeast artificial chromosome (YAC) clones along a high-resolution molecular linkage map carrying 118 DNA markers distributed over 123.7 cM of genomic DNA. YAC clones have been identified by colony and Southern hybridization for 105 restriction fragment length polymorphism (RFLP) markers and by polymerase chain reaction (PCR) screening for 8 sequence-tagged site (STS) markers and 5 randomly amplified polymorphic DNA (RAPD) markers. Of 458 YACs, 235 individual YACs with an average insert length of 350 kb were selected and ordered on chromosome 5 from the YAC library. Forty-eight contigs covering nearly 21 Mb were formed on the chromosome 5; the longest one was 6 cM and covered 1.5 Mb. The length covered with YAC clones corresponded to 62% of the total length, of chromosome 5. There were many multicopy sequences of expressed genes on chromosome 5. The distribution of many copies of these expressed gene sequences was determined by YAC Southern hybridization and is discussed. A physical map with these characteristics provides a powerful tool for elucidation of genome structure and extraction of useful genetic information in rice.

摘要

利用酵母人工染色体(YAC)克隆构建了水稻第5号染色体的物理图谱,该图谱沿着一个高分辨率分子连锁图谱构建,该连锁图谱携带118个DNA标记,分布在123.7厘摩的基因组DNA上。通过菌落和Southern杂交鉴定YAC克隆,用于105个限制性片段长度多态性(RFLP)标记,通过聚合酶链反应(PCR)筛选用于8个序列标签位点(STS)标记和5个随机扩增多态性DNA(RAPD)标记。在458个YAC中,选择了235个平均插入长度为350 kb的单个YAC,并从YAC文库中将其排列在第5号染色体上。在第5号染色体上形成了48个覆盖近21兆碱基的重叠群;最长的一个是厘摩,覆盖1.5兆碱基。YAC克隆覆盖的长度相当于第5号染色体总长度的62%。第5号染色体上有许多表达基因的多拷贝序列。通过YAC Southern杂交确定了这些表达基因序列多拷贝的分布,并进行了讨论。具有这些特征的物理图谱为阐明水稻基因组结构和提取有用的遗传信息提供了有力工具。

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