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SSH法获取水稻矮化突变体相关的cDNA片段

Identification of cDNA Fragments Differentially Expressed Between a Rice Mutant dwarf69 and Wild Type TGA-2 by Suppressive Subtraction Hybridization

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【作者】 王永胜王景李发强刘筱斌刘良式

【Author】 Wang Yongsheng, Wang Jing, Li Faqiang, Liu Xiaobin , Liu Liangshi (The Key Laboratory of Gene Engineering of Ministry of Education, Zhongshan University, Guangzhou 510275) ( South China Institute of Botany, Chinese Academic Sciences, Guangzhou 51

【机构】 中山大学基因工程教育部重点实验室!广州51027中科院华南植物研究所!广州510600

【摘要】 利用抑制性消减杂交 (Suppressivesubtractionhybridization ,SSH)分离水稻矮化突变体dwarf69与其野生型对照品系特光矮 2 (Oryzasativa ,TGA 2indica)间表达有差异的cDNA片段。分别以dwarf69作为驱赶子 ,TGA 2为检测子 ,以及以dwarf69作为检测子 ,TGA 2为驱赶子建立了两个差异表达cDNA文库 ,分别代表在TGA 2和dwarf69特异表达的cDNA。经反式Northern检测这两个文库共得到 1 0个阳性克隆。序列分析表明有两个序列为水稻中首次报道 ,一个序列有相应的基因组序列报道 ,但其cDNA序列是首次报道。

【Abstract】 The suppressive subtraction hybridization (SSH) technique was used to isolate the cDNA fragments that showed differential expression between the rice mutant dwarf69 and wild type strain,TGA 2(( Oryza sativa, indica).Two differential expressing cDNA libraries were constructed using the cDNAs of dwarf69 as driver and the cDNAs of TGA 2 as tester, vice versa. These two cDNA libraries indicated the differential expressing cDNA fragments in TGA 2 and dwarf69. By reverse Northern blots, ten positive clones were obtained. Subsequently sequencing analysis showed two cDNA fragments were first reported in rice, and another was reported as genomic sequence previously but not as cDNA.

【基金】 教育部重点科学技术 ( 0 61C615 );广东省科技计划 ( 99M 0 15 0 3G)资助项目
  • 【文献出处】 高技术通讯 ,High Technology Letters , 编辑部邮箱 ,2001年05期
  • 【分类号】Q943
  • 【被引频次】41
  • 【下载频次】195
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