水稻冷胁迫诱导的甲基化差异片段CIDM7的分离和分析  

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

作　　者：华扬[1] 陈学峰[1] 熊建华[1] 张义平[1] 朱英国[1] 

机构地区：[1]武汉大学植物发育生物学教育部重点实验室,武汉430072

出　　处：《遗传》2005年第4期595-600,共6页Hereditas（Beijing)

基　　金：国家"973"项目(编号:2001CB108805)~~

摘　　要：采用基于AFLP的甲基化敏感扩增多态性的方法对5℃冷处理48h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48h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(coldinducingdifferentialmethylation)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1422bp,编码425个氨基酸,经氨基酸序列分析为一个具有Fbox结构的假定蛋白。CIDM7为单拷贝,定位于日本晴10号染色体12.56～12.57Mb。By using restriction enzyme isoschizomers that differ in their sensitivity to methylation of their recognition sites, AFLP analysis was 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 were found in some CCGG sites. Some differentially-methylated encoding sequences were isolated, of which a fragment named CIDM7 （cold-inducing differential methylation） was sequenced and executed BLAST against Nipponbare cDNA data, thus we obtained its full length cDNA of 1422 bp encoding a hypothetical F-box protein （425 aa）. CIDM7 was demethylated induced by cold stress. Northern blot analysis confirmed that CIDM7 gene expression was up-regulated by cold stress. CIDM7 is single copy and lacated on the Nipponbare chromosome 10 （12.56-12.57 Mb）.

关 键 词：胞嘧啶甲基化 水稻 AFLP 冷胁迫 

分 类 号：S511[农业科学—作物学]