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.
出处
《分子植物育种》
CAS
CSCD
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。
关键词
水稻
SMART技术
干旱胁迫
全长CDNA文库
抗旱相关基因
Rice （Oryza sativa L.）, SMART technique, Drought stress, Full-length eDNA library, Drought-tolerance related genes
