玉米醇溶蛋白γ-zein基因载体构建及亚细胞定位  

Construction and Subcellular Localization of Plant Expression Vector with Prolamine Protein from Corn and GFP Gene

作　　者：张玉[1] 白史且[1] 李聪[2] 李达旭[1] 邓永昌[1] 王涌鑫[2] 

机构地区：[1]四川省草原科学研究院,成都犀浦611731 [2]中国农业科学院北京畜牧兽医研究所,北京100094

出　　处：《分子植物育种》2011年第2期174-179,共6页Molecular Plant Breeding

基　　金：国家十二五科技支撑计划(2011BAD17B03);四川省应用基础项目(2008JY0010);四川省科技厅十一五牧草育种攻关(06SG023-001);国家牧草产业技术体系阿坝综合试验站共同资助

摘　　要：用PCR方法从pROK.TG1LK扩增得到γ-zein基因的全长,克隆到中间载体pMD18-T,获得了重组质粒pMD18-γ-zein,分别用NcoⅠ和BglⅡ2种限制性内切酶酶切重组质粒和经过改良的pCAMBIAI1302载体,经回收、连接、转化、鉴定后,构建了由35S驱动的绿色荧光蛋白(GFP)融合的植物表达载体pCB-GFP-γ-zein,用基因枪法将GFP-γ-zein基因整合到洋葱表皮细胞中,通过共聚焦显微镜观察,该融合基因在洋葱表皮细胞的细胞壁、细胞核和细胞质中得到了表达。The total length sequence of γ-zein gene in pROK.TG1LK plasmid was amplified by PCR. The fragment was cloned into pMD18-T middle vector, and a new recombined vector named pMD18-T-zein was obtained. A new plant expression vector named pCB-GFP-γ-zein in which the reporter gene GFP is droved by 35s was constructed after cutting two vector pMD18-γ-zein and pCAMBIAI1302 with NcoⅠ and BglⅡ restriction enzymes, subsequently reclamation, ligation, transformation and identification. The recombined plant expression vector was incorporated into epidermic cell of onion by gene gun method and the fusion gene was transient expressed. The result of transient expression showed that the gene located in cell wall, cell, nucleus and cytoplasm.

关 键 词：绿色荧光蛋白(GFP) γ-zein基因 植物表达载体 亚细胞定位 

分 类 号：S513[农业科学—作物学]