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水稻种子萌发期高压诱导的SSH-cDNA文库构建与分析
引用本文:刘勋成,张美,段俊.水稻种子萌发期高压诱导的SSH-cDNA文库构建与分析[J].高压物理学报,2008,22(4).
作者姓名:刘勋成  张美  段俊
作者单位:1. 中国科学院华南植物园,广东广州,510650;中国科学院研究生院,北京,100039
2. 中国科学院华南植物园,广东广州,510650
基金项目:广东省自然科学基金重点项目(05103637);;华南植物园科研启动经费(0553011001);;广东省科技攻关计划项目(2006B20201016)
摘    要: 为了研究高压处理对高等植物基因表达的影响,以萌发过程中经高压处理后的水稻种子的mRNA作为检测子(Tester),以对照样品的mRNA作为驱动子(Driver),采用抑制差减杂交(SSH)结合镜像选择技术(MOS)构建了高压处理后水稻种子萌发过程中诱导表达的差减cDNA文库,并通过蓝白斑筛选获得了580个重组克隆。菌液PCR检测的结果表明这些克隆中的插入片段的大小主要分布在200~1000 bp之间,平均大小约为575 bp,其中单一扩增条带的有效重组克隆数为542个,有效的重组克隆率为93.3%。对部分重组克隆进行测序发现,受高压诱导表达的基因主要涉及到代谢、胁迫响应、转录调控等方面。为高压处理后水稻种子基因表达模式变化的研究打下了基础。

关 键 词:高压  抑制差减杂交  水稻
收稿时间:2007-11-28;

Construction and Analysis of SSH-cDNA Library Induced by High Pressure in Rice Seeds during Germination
LIU Xun-Cheng,ZHANG Mei,DUAN Jun.Construction and Analysis of SSH-cDNA Library Induced by High Pressure in Rice Seeds during Germination[J].Chinese Journal of High Pressure Physics,2008,22(4).
Authors:LIU Xun-Cheng  ZHANG Mei  DUAN Jun
Institution:1.South China Botanic Garden;Chinese Academy of Sciences;Guangzhou 510650;China;2.Graduate University of Chinese Academy of Sciences;Beijing 100039;China
Abstract:In order to study the effect of expression pattern of high hydrostatic pressure on high plant,a SSH-cDNA library was constructed with rice germinating seeds in this paper.Suppression subtractive hybridization(SSH) in combination with mirror orientation selection(MOS) was performed using the mRNA from rice germinating seeds treated by 75 MPa high hydrostatic pressure as tester and the mRNA from control(seeds untreated by high hydrostatic pressure) as the driver.580 recombinant clones were picked through blue...
Keywords:high hydrostatic pressure  suppression subtractive hybridization (SSH  rice
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