| 绵毛优若藜冷诱导SSH文库构建研究 |
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| 引用本文： | 张党权,明付焕,江平,樊绍刚,杨自辉,邓顺阳.绵毛优若藜冷诱导SSH文库构建研究[J].中南林业科技大学学报,2010,30(2). |
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| 作者姓名： | 张党权 明付焕 江平 樊绍刚 杨自辉 邓顺阳 |
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| 作者单位： | 张党权,明付焕,樊绍刚,邓顺阳(中南林业科技大学,经济林育种与栽培国家林业局重点实验室,湖南,长沙,410004);江平(兰州大学,土木工程与力学学院,甘肃,兰州,730000);杨自辉(甘肃省治沙研究所,甘肃省荒漠化防治重点实验室,甘肃,武威,733000) |
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| 基金项目： | 国家自然科学基金资助项目，中南林业科技大学青年基金重点资助项目 |
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| 摘 要： | 绵毛优若藜是我国近年成功引种的高抗逆性饲用及防风固沙植物。以常温下的绵毛优若藜叶片和嫩枝为对照组,冷胁迫诱导处理后的叶片和嫩枝为试验组,获得两者高质量mRNA后反转录成cDNA,RsaI消化使其平末端化,试验组cDNA的5’末端连接上2个特异接头,然后分别与过量对照组cDNA进行杂交,把两份杂交样品混合后,与过量的新鲜变性的对照组cDNA过夜杂交,杂交后的混合液稀释后进行第一轮抑制性PCR以选择扩增特异表达的cDNA,然后以第一轮PCR产物为模板用巢式引物进行二次抑制性PCR,获得的富集差异表达cDNA即为绵毛优若藜冷胁迫SSH文库。将SSH文库的cDNA插入到载体中,共获得了362个克隆,为后续绵毛优若藜的抗冻基因克隆与转基因应用奠定了基础。
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| 关 键 词： | 绵毛优若藜 抑制消减杂交文库 冷胁迫 抗冻基因 cDNA克隆 |
Construction of cold-stressed SSH library of Ceratoides lanata and sequence analysis of its differentially expressed cDNA |
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| ZHANG Dang-quan,MING Fu-huan,JIANG Ping,FANG Shao-gang,YANG Zhi-hui,DENG Shun-yang.Construction of cold-stressed SSH library of Ceratoides lanata and sequence analysis of its differentially expressed cDNA[J].Journal of Central South Forestry University,2010,30(2). |
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| Authors: | ZHANG Dang-quan MING Fu-huan JIANG Ping FANG Shao-gang YANG Zhi-hui DENG Shun-yang |
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| Institution: | ZHANG Dang-quan1,MING Fu-huan1,JIANG Ping2,FANG Shao-gang1,YANG Zhi-hui3,DENG Shun-yang1(1.The Key Lab of Non-wood Forest Products of State Forestry Administration,Central South University of Forestry , Technology,Changsha 410004,Hunan,China,2.School of Civil Engineering , Mechanics,Lanzhou University,Lanzhou 730000,Gansu,3.The Key Lab of Desertification Control,Gansu Desert Control Research Institute,Wuwei 733000,China) |
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| Abstract: | As a high stress resistance psammophyte with the characteristic of windbreak and sand fixation,Ceratoides lanata had been successfully introduced into China in recent years.The leaves and twigs of Ceratoides lanata in room temperature were used as the driver and those treated by cold stress were used as tester.The tester and driver ds cDNA were prepared from their high quality mRNA which were purified from their total RNA.The tester and driver cDNA were separately digested to obtain shorter,blunt-ended mole... |
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| Keywords: | Ceratoides lanata SSH library cold stress antifreeze gene cDNA cloning |
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