盐胁迫下短芒大麦cDNA文库的构建  

Construction of cDNA Library of Hordeum brevisubulatium Link under Salt Stress

作　　者：陆一鸣[1] 李彦舫[1] 白杰英[1] 曹明富[2] 马鹤雯[1] 阮承迈[3] 

机构地区：[1]解放军军需大学生物教研室,吉林长春130062 [2]杭州师范大学生物系,浙江杭州3100363 [3]军事医学科学院,北京100850

出　　处：《吉林农业大学学报》2003年第5期499-502,共4页Journal of Jilin Agricultural University

基　　金：国家自然科学基金(39800106);吉林省自然科学基金(970205-2)

摘　　要：为了克隆与研究盐胁迫相关的基因,以盐胁迫后的短芒大麦叶片为材料,用Trizol一步法提取总RNA,Oligo(dT)纤维素柱纯化mRNA,反转录合成第一链cDNA和第二链cDNA.cDNA纯化后,与λZAP表达载体连接,体外包装后得到盐胁迫下短芒大麦cDNA文库,其重组率达96%,滴度为2×107 pfu/mL,插入片段长度0.5～3.0 kb.In order to clone and study salt-stress related genes, total RNA was-isolated from leaves ofHordeum brevisubulatium(Trin. )Link using TRIZOL reagent under salt stress and mRNA was further purified through Oligo-dT cellulose. The first strand cDNA was synthesized by reverse transcriptase. After the synthesis of the second strand, the cDNA was purified and ligated with λZAP express vector. After packaging in vitro, the cDNA library of Hordeum brevisubulatium(Trin. )Link was constructed. The cD-NA library contained 2×10~7 clones, in which more than 96% clones were recombinants and most of in- sert DNA fragments were between 0.5～3.0 kb.

关 键 词：盐胁迫 短芒大麦 CDNA文库 Trizol一步法 提取 纯化 

分 类 号：S512.3[农业科学—作物学]