百脉根液泡膜H^+-PPase基因LcVP1植物表达载体构建与转基因毛白杨的获得  

Construction of Plant Expression Vector of Tonoplast H^+-PPase from Lotus corniculatus L.and Obtaintion Transgenic Populus tomentosa Carr.

作　　者：孙艳香[1] 冯雪[1] 赵学良[1] 

机构地区：[1]廊坊师范学院生命科学学院遗传与育种研究所,河北廊坊065000

出　　处：《中国农学通报》2011年第4期134-138,共5页Chinese Agricultural Science Bulletin

基　　金：河北省教育厅自然科学项目"豆科模式植物百脉根液泡膜H+-PPase基因在植物抗旱耐盐基因工程中的应用研究"(20070318);廊坊市科技攻关项目"利用基因工程手段选育抗旱;耐盐林木新品系"(2007010201)

摘　　要：为通过基因工程手段进行杨树耐盐性状的遗传改良,笔者分析已克隆的百脉根液泡膜H+-PPase基因LcVP1的cDNA序列,根据其与植物表达中间载体pGN上的酶切位点设计1对带酶切位点的特异性引物,以测序质粒为模板,PCR扩增百脉根液泡膜H+-PPase基因开放阅读框片段。双酶切PCR回收产物和pGN载体,将目的片段定向连接构建成植物表达载体pGVP,并转入根癌农杆菌;在此基础上,利用根癌农杆菌介导的叶盘转化法,将百脉根液泡膜H+-PPase基因转入毛白杨基因组中。转基因杨树的获得,为获得耐盐性提高的杨树品系提供了基础。In order to improve salt tolerance ability of poplar,in the present study,according to the restrictio nenzyme site of pGN vector and the tonoplast H＋-PPase gene LcVP1 cDNA sequence from Lotus corniculatus L.,a pair of primers containing restriction enzyme site was desigined and used to amplify LcVP1 ORF.PCR pruducts and the plasmid pGN were digested by the corresponding restricted enzymes respectively,and linked directionally.Constructed plasmid was named pGVP,and transformated into Agrobacterium.Then,Agrobacterium-mediated leaf discs genetic transformation was carried out,and LcVP1 had been integrated into the genome of the transgenic Populus tomentosa Carr.

关 键 词：百脉根 液泡膜H+-PPase LcVP1 pGVP 毛白杨 

分 类 号：S3[农业科学—农艺学]