摘要
应用PrimerPremier5.0软件,根据GenBank数据库报道的查尔酮合成酶基因启动子序列(EF199747)设计1对特异性PCR扩增引物,以矮牵牛品种‘午夜蓝色’叶片总DNA为模板,用TaqDNA聚合酶成功扩增出1条约0.5kb的DNA片段,回收该片段并连接到pMD18-T载体上。结果表明:经测序该启动子片段长550bp;bl2seq分析结果表明该启动子与目标序列相似性高达100%;PLACE在线分析显示在克隆片段中含有TATAbox、CAATbox、capsite、antherbox、box1、box2、Gbox及TACPyAT-box等顺式元件;并构建了矮牵牛CHS基因启动子融合标记基因GUS的植物表达载体pPhCHS::GUS。
Based on the Primer Premier 5.0software,apair of specific PCR primers was designed according to promoter sequence of CHS gene of Petunia hybridareported in GenBank（EF199747）.A DNA fragment about 0.5kb was successfully amplified with the genome DNA of Petunia hybridcultivar＇Midnight＇as template and Taq polymerase as DNA polymerase.The purified fragment of PCR products was ligated with pMD18-T vector,and the sequencing result showed that the length of the promoter of CHS gene of Petuniah ybrid was 550bp.Bl2seq analysis revealed that the sequence similarity between the cloned promoter sequence and EF199747was up to 100%.Online PLACE analysis indicated that it contained cis-elements such as TATA-box,CAAT-box,cap site,anther-box,box1,box2,G-box and TACPyAT-box.And plant expression vector of pPhCHS：：GUSthat fused the promoter of CHSgene of Petuniah ybridaand marker gene GUSwas successfully constructed.
出处
《北方园艺》
CAS
北大核心
2010年第17期155-158,共4页
Northern Horticulture
基金
国家自然科学基金资助项目(30740013)
林木
花卉遗传育种教育部重点实验室开放基金资助项目(05-03)
关键词
矮牵牛
CHS基因启动子
克隆
序列分析
植物表达载体
Petunia hybrid
promoter of CHSgene
cloning
sequence analysis
plant expression vector
