干旱胁迫下耐旱稻中旱3号全长cDNA文库的构建  

Construction of Full Length cDNA Library from Drought-tolerant Rice cv. Zhonghan No. 3 Cultivated under Drought Stress

作　　者：刘运华[1,2] 刘灶长[1] 周立国[1,2] 余舜武[1] 刘鸿艳[1] 罗利军[1] 

机构地区：[1]上海市农业生物基因中心 [2]华中农业大学植物科学院,武汉430070

出　　处：《分子植物育种》2007年第3期367-370,共4页Molecular Plant Breeding

基　　金：上海市科委重大基础研究项目(03DJ14015);上海市浦江人才计划(05PJ14085)资助。

摘　　要：干旱胁迫导致很多基因的表达发生变化。以干旱胁迫下的耐旱稻品种中旱3号为材料,我们用SMART技术（RNA转录5′末端模板转换技术）成功构建一个高质量的干旱胁迫诱导的节水抗旱稻中旱3号全长cDNA文库。体外包装后感染大肠杆菌XL1-Blue宿主菌,未扩展文库的滴度为1.94×10^6pfu/mL,重组率为85.15%。扩展后文库的滴度为1.45×10^11pfu/mL。将λ噬菌体臂转入pTriplEx2质粒,从中随机挑选100个克隆,PCR扩增检测插入片段长度,结果显示文库插入片段在500～2000bp之间。Drought stress affects plant physiological process by causing altered gene expression. A high quality full-length cDNA library of drought-stressed rice cv. Zhonghan 3 was constructed with SMART （switching mechanism at 5′ end of the RNA transcript） technique. The λ phage packaging reaction and library amplification in E.coli strain XL1-Blue were performed. The titer of unamplified library was 1.94× 10^6 pfu/mL and the recombinant rate was 85.15%. The amplified library obtained a 1.45× 10^11 pfu/mL recombinants. Finally the λTriplEx2 clone was transformed to pTriplEx2 plasmid and 100 clones were picked randomly from the library for screening size of cDNA inserts through PCR. Results indicated that most of the inserts were between 500 and 2 000 bp.

关 键 词：水稻 SMART技术 干旱胁迫 全长CDNA文库 抗旱相关基因 

分 类 号：S511[农业科学—作物学]