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用于水稻基因图位克隆的全基因组DNA多态性数据库的开发。

Development of genome-wide DNA polymorphism database for map-based cloning of rice genes.

作者信息

Shen Ying-Jia, Jiang Hua, Jin Jian-Peng, Zhang Zai-Bao, Xi Biao, He You-Yu, Wang Guan, Wang Chen, Qian Lily, Li Xiang, Yu Qing-Bo, Liu Hui-Juan, Chen De-Hui, Gao Jian-Hua, Huang Hai, Shi Tie-Liu, Yang Zhong-Nan

机构信息

College of Life and Environment Sciences, Shanghai Normal University, Shanghai 200234, China.

出版信息

Plant Physiol. 2004 Jul;135(3):1198-205. doi: 10.1104/pp.103.038463.

Abstract

DNA polymorphism is the basis to develop molecular markers that are widely used in genetic mapping today. A genome-wide rice (Oryza sativa) DNA polymorphism database has been constructed in this work using the genomes of Nipponbare, a cultivar of japonica, and 93-11, a cultivar of indica. This database contains 1,703,176 single nucleotide polymorphisms (SNPs) and 479,406 Insertion/Deletions (InDels), approximately one SNP every 268 bp and one InDel every 953 bp in rice genome. Both SNPs and InDels in the database were experimentally validated. Of 109 randomly selected SNPs, 107 SNPs (98.2%) are accurate. PCR analysis indicated that 90% (97 of 108) of InDels in the database could be used as molecular markers, and 68% to 89% of the 97 InDel markers have polymorphisms between other indica cultivars (Guang-lu-ai 4 and Long-te-pu B) and japonica cultivars (Zhong-hua 11 and 9522). This suggests that this database can be used not only for Nipponbare and 93-11, but also for other japonica and indica cultivars. While validating InDel polymorphisms in the database, a set of InDel markers with each chromosome 3 to 5 marker was developed. These markers are inexpensive and easy to use, and can be used for any combination of japonica and indica cultivars used in this work. This rice DNA polymorphism database will be a valuable resource and important tool for map-based cloning of rice gene, as well as in other various research on rice (http://shenghuan.shnu.edu.cn/ricemarker).

摘要

DNA多态性是开发分子标记的基础,这些分子标记如今广泛应用于基因图谱绘制。在本研究中,利用粳稻品种日本晴和籼稻品种93-11的基因组构建了一个全基因组水稻(Oryza sativa)DNA多态性数据库。该数据库包含1,703,176个单核苷酸多态性(SNP)和479,406个插入/缺失(InDel),在水稻基因组中大约每268 bp有一个SNP,每953 bp有一个InDel。数据库中的SNP和InDel均经过实验验证。在随机选择的109个SNP中,107个SNP(98.2%)是准确的。PCR分析表明,数据库中90%(108个中的97个)的InDel可作为分子标记,并且这97个InDel标记中有68%至89%在其他籼稻品种(广陆矮4号和龙特甫B)和粳稻品种(中华11号和9522)之间具有多态性。这表明该数据库不仅可用于日本晴和93-11,还可用于其他粳稻和籼稻品种。在验证数据库中InDel多态性的同时,开发了一组每条染色体有3至5个标记的InDel标记。这些标记成本低廉且易于使用,可用于本研究中使用的任何粳稻和籼稻品种组合。这个水稻DNA多态性数据库将成为基于图谱克隆水稻基因以及水稻其他各种研究的宝贵资源和重要工具(http://shenghuan.shnu.edu.cn/ricemarker)。

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