摘要
从小麦中分别克隆了包括rbcL基因 3′端部分和完整的 psaI、ycf4基因的DNA片段。利用克隆到的DNA片段作为同源重组片段、烟草叶绿体 16SrRNA基因的启动子Prrn和PsbA基因的终止子PsbA3′控制筛选标记基因aadA和报告基因gfp的转录 ,构建了小麦叶绿体基因组定点整合表达载体pRAGY。用该载体转化大肠杆菌 ,在激光扫描共聚焦显微镜下 ,检测到了gfp基因成功表达的产物被激发出的强烈的绿色荧光。
A site specific integration and expression vector for wheat chloroplast genetic transformation (Fig.3) was constructed for the first time. It contains the homologous recombinant fragment 1, the 3′end part of rbcL and the homologous recombinant fragment 2,a DNA fragment embracing the two gene: psaI and ycf4 . the homologous recombinant fragments were amplified from the genome of wheat; the selectable marker aadA gene and the reporter gene gfp were regulated by the promoter Prrn and the terminator PsbA3′from tobacco. When the E. coli was transformed by the expression vector, intensive green fluorescence produced by the products of gfp gene was observed through laser co focal microscope (Fig.4). This result indicate that the expression vector we have constructed here can be promisingly used in prokaryotic chloroplast transformation of wheat and the foreign gene in it can express successfully.
出处
《植物生理与分子生物学学报》
CAS
CSCD
2003年第5期469-473,共5页
Journal Of Plant Physiology and Molecular Biology
基金
国家自然科学基金项目 (NO .3 0 2 70 82 9)
山东省科技计划专项(NO .0 12 10 0 10 4)资助
关键词
小麦
叶绿体基因组
表达载体构建
GFP基因
定点整合
wheat
chloroplast genome
construction of expression vector
gfp gene
site specific integration
作者简介
侯丙凯,通讯作者(E-mail:bkhou@sdu.edu.cn;Tel:0531-8364525).
