Construction of cDNA Library from Populus euphratica

编号 zgly0000301815

文献类型 期刊论文

文献题名 Construction of cDNA Library from Populus euphratica

学科分类 220.99;林学其他学科

作者 YuGuangjun  WangYiqin  ShenXin 

作者单位 CollegeofBiologicalSciencesandBiotechnology 

母体文献 中国林学: 英文版 

年卷期 2003,5(2)

页码 7-9

年份 2003 

分类号 S792.11 

关键词 杨树  cDNA文库  耐盐基因  盐胁迫  磁性有孔小珠 

文摘内容 In order to isolate and clone salt-tolerance involved genes of Populus euphratica, we constructed a cDNA library from salt-treated leaves of P. euphratica. In the experiment, double strand cDNA were synthesized by a beads-based method. The syntheses of the first strand and the second strand cDNA, adapter ligation and restriction reaction for releasing cDNA were all conducted on thc beads. The double strand cDNA werc released from magnetic beads by digestion with Notl, and cDNA fragments smaller than 500 bp and residual adapters were removed through cDNA size fractionation columns. Finally, double strand cDNA wcre directionally cloned into λExcell vector. The results show that the primary titer of the cDNA library is 7.46 × 10^6 pfu per mL and the packaging efficiency reaches 1.47 × 10^7 recombinants per μg DNA. λ DNA extracted from two clones of plaque were digested by EcoR 1 and Notl, both of the clones contained inserts larger than 900 bp. Thcse results show that the cDNA library of salt-treated P. euphratica leaves has been successfully constructed。