遗传 ›› 2005, Vol. 27 ›› Issue (4): 595-600.

• 研究报告 • 上一篇    下一篇

水稻冷胁迫诱导的甲基化差异片段CIDM7的分离和分析

华 扬;陈学峰;熊建华;张义平;朱英国   

  1. 武汉大学发育生物学教育部重点实验室,武汉430072
  • 收稿日期:2004-06-02 修回日期:2005-02-21 出版日期:2005-08-10 发布日期:2005-08-10
  • 通讯作者: 华扬

Isolation and analysis of Isolation and Analysis of Differentially-Methylated Fragment CIDM7 in Rice Induced by Cold Stress

HHA Yang, CHEN Xue-Feng, XIONG Jian-Hua, ZHANG Yi-Ping, ZHU Ying-Guo   

  1. Key Laboratory of MOE forDevelopmental Biology, Wuhan University, Wuhan 430072, China
  • Received:2004-06-02 Revised:2005-02-21 Online:2005-08-10 Published:2005-08-10
  • Contact: Yang Hua

摘要: 文章采用基于AFLP的甲基化敏感扩增多态性的方法对5℃冷处理48小时的水稻9311叶片DNA胞嘧啶甲基化模式进行了初步研究,发现冷处理48小时后9311基因组中一些CCGG位点发生了重新甲基化或去甲基化,并获得一些与水稻cDNA高度同源的甲基化差异片段,其中CIDM7 (cold-inducing differential methylation)片段在冷胁迫后去甲基化。Northern杂交证明CIDM7在冷胁迫后增强表达。通过在日本晴全长cDNA数据库中的同源搜索获得其全长cDNA序列,该cDNA序列全长1422 bp,编码425个氨基酸,经氨基酸序列分析为一个具有F-box结构的假定蛋白。CIDM7为单拷贝,定位于日本晴10号染色体12.56 ~12.57 Mb。

关键词: 水稻, 胞嘧啶甲基化, 冷胁迫, AFLP

Abstract: By using restriction enzyme isoschizomers that differ in their sensitivity to methylation of their recognition sites, AFLP analysis is carried out to analyse differences of the methylation state of anonymous CCGG sequences between cold-treated(5℃ for 2 d) and control 9311(Oryza sativa L.) leaves DNA.Demethylation or De novo methylation induced by cold stress are found in some CCGG sites. Some differentially-methylated encoding sequences are isolated, of which a fragment named CIDM7 (cold-inducing differential methylation) is sequenced and executed BLAST against Nipponbare cDNA data, thus we obtain its full length cDNA of 1422 bp encoding a hypothetical F-box protein(425 aa).CIDM7 is demethylated induced by cold stress. Northern blot analysis confirms that CIDM7 gene expression is up-regulated by cold stress. CIDM7 is single copy and lacated on the Nipponbare chromosome 10 (12.56 Mb~12.57 Mb).

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