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用抑制差减杂交法分离小麦幼苗水分胁迫诱导表达的cDNA

Isolation and Cloning of cDNA Induced by Water Stress in Wheat Seedling Using Suppression Subtractive Hybridization

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【作者】 王转贾晋平景蕊莲

【Author】 Wang Zhuan 1 Jia Jinping 2 Jing Ruilian 1 ( 1 Key Laboratory of Crop Germplasm & Biotechnology Ministry of Agriculture Institute of Crop Germplasm Resources Chinese Academy of Agricultural Sciences, Beijing 100081; 2 Chinese Agricultural University, Beijing 100094)

【机构】 中国农业科学院作物品种资源研究所农业部作物种质资源与生物技术重点实验室中国农业大学生物学院中国农业科学院作物品种资源研究所农业部作物种质资源与生物技术重点实验室 北京100081北京100094北京100081

【摘要】 小麦种子水培 ,幼苗长至一叶一心后 ,在 2 0℃生长箱中水培 4 8h作为对照组 (Driver) ,PEG 6 0 0 0水溶液胁迫培养 4 8h作为处理组 (Tester)。进行抑制差减杂交 ,构建包含 15 0 0个独立克隆的SSH文库。以正向和反向差减杂交后的cDNA为探针 ,筛选SSH文库 ,得到 181个阳性克隆 ,测序后获不重复EST 10 1个。

【Abstract】 Hanxuan No.10 wheat (Triticum aestivum L.) seedlings were cultivated in water at 20℃ with 12h light every day until one and a half leaves. Then half seedlings were continuously cultivated in water for 48h as control (the driver), another half seedlings were stressed in PEG 6000(-0.5MPa)solution for 48h as treatment (the tester). A subtracted cDNA library was constructed by suppression subtractive hybridization (SSH), which was composed of approximately 1500 individual recombinant clones. A total of 181 positive clones were obtained by screening the library using forward and reverse subtracted cDNA probes, and 101 unique ESTs induced by water stress were gained based on the sequencing data.

【基金】 国家“8 63”项目 ( 2 0 0 1AA 2 110 5 1)资助
  • 【文献出处】 生物技术通报 ,Biotechnology Information , 编辑部邮箱 ,2003年05期
  • 【分类号】S512.1
  • 【被引频次】75
  • 【下载频次】329
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