| 曹玲珑，李冬兵，熊大斌，牛洪斌，姜玉梅，尹 钧.外源ABA处理大麦胚的均一化cDNA文库的构建[J].麦类作物学报,2014,34(7):912 |
| 外源ABA处理大麦胚的均一化cDNA文库的构建 |
| Construction of a Normalized cDNA Library of Barley Embryo Treated with Exogenous ABA |
| |
| DOI：10.7606/j.issn.1009-1041.2014.07.06 |
| 中文关键词: 外源ABA 大麦 cDNA文库 |
| 英文关键词:Exogenous ABA Barley cDNA library |
| 基金项目:国家转基因新品种培育重大专项（2011ZX08002 003）；作物分子育种与品种创制项目（2012AA101105） |
|
| 摘要点击次数: 1006 |
| 全文下载次数: 833 |
| 中文摘要: |
| 构建外源ABA处理大麦胚的均一化cDNA文库是利用酵母双杂交方法筛选与靶蛋白相互作用蛋白的前提条件。为给大麦抗逆基因的研究及抗逆转基因大麦材料的创制奠定基础，利用5 μmol·L-1 ABA和蒸馏水分别处理大麦种子0 h、6 h、12 h、24 h，剥取大麦的胚，用热酚法提取总RNA，利用SMART原理进行反转录PCR并进行均一化处理获得ds cDNA。经SfiⅠ酶切的ds cDNA与pGADT7 Rec载体连接后，转化大肠杆菌感受态细胞DH10B构建文库。结果表明，文库的容量为1.1×106，插入片段大于1 000 bp，集中在1 000～3 000 bp之间，符合酵母双杂交文库的要求。 |
| 英文摘要: |
| It is necessary to construct the cDNA library of barley embryo before screening the proteins interacted with target proteins．In the experiment reported in this paper,barley seeds were treated with 5 μmol·L-1 ABA and distilled water for 0 h,6 h,12 h and 24 h,respectively．Barley embryos were stripped and total RNA was extracted. Ds cDNA was synthesized by RT PCR with SMART method．The ds cDNA which digested by Sfi I was ligated to the vectors pGADT7 Rec．All the recombinant vectors were electroporated into DH10B to construct the cDNA library．The detection results showed that the cDNA library contained 1.1×106 recombinant clones and that all the inserted cDNA fragments were more than 1 kb in size,between 1 kb and 3 kb and most were around 1.2 kb,which meets the requirement of the yeast two hybrid library. |
|
查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
基本信息
