水稻种子萌发期高压诱导的SSH-cDNA文库构建与分析

刘勋成; 张美; 段俊

doi:10.11858/gywlxb.2008.04.006

水稻种子萌发期高压诱导的SSH-cDNA文库构建与分析

doi: 10.11858/gywlxb.2008.04.006

刘勋成^1,2,
张美¹,
段俊^1,

1.
中国科学院华南植物园，广东广州 510650；
2.
中国科学院研究生院，北京 100039

详细信息

通讯作者:
段俊

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出版历程
- 收稿日期: 2007-11-28
- 修回日期: 2008-03-28
- 发布日期: 2008-12-05

Construction and Analysis of SSH-cDNA Library Induced by High Pressure in Rice Seeds during Germination

1.
South China Botanic Garden, Chinese Academy of Sciences, Guangzhou 510650, China;
2.
Graduate University of Chinese Academy of Sciences, Beijing 100039, China

More Information

Corresponding author: DUAN Jun

摘要

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。
- 高压 /
- 抑制差减杂交 /
- 水稻
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/white screening. PCR detection of the recombinant clones showed that the size of inserts mainly ranged from 200 bp to 1000 bp, and the average size of these inserts were about 575 bp. Among the 580 recombinant clones, 542 clones (93.3%) were identified to be effective single band ones. After sequencing some of the recombinant clones, it was found that the genes induced by high hydrostatic pressure were mainly involved in metabolism, stress response and transcriptional regulation. The research paved the way for studying the gene expression patterns of rice seeds responding to high hydrostatic pressure.
- high hydrostatic pressure /
- suppression subtractive hybridization (SSH) /
- rice

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参考文献(21)

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