豆荚特异性启动子Pmsg的克隆及抗大豆食心虫植物表达载体的构建

植物基因工程载体构建时,一般采用组成型启动子,因涉及转基因食品安全性问题,在社会上引起争议。采用组织特异性启动子代替组成型启动子,既可调控下游基因的表达又能解决转基因食品安全性问题。采用同源克隆方法从栽培大豆绥农14中克隆了2 278 bp的豆荚特异性启动子Pmsg,与GenBank上的大豆msg基因启动子序列相似性为99%。构建了由启动子Pmsg调控经人工改造、具有抗大豆食心虫功能的基因(Cry1Iem)的表达载体pBMBt,以及由种子特异性启动子PGy2调控Cry1Iem基因的表达载体pBGBt,为抗大豆食心虫基因工程研究奠定了重要基础。

Clone of the pod-specific promoter and construction of expression vectors resistant to soybean bean pod borer

The constitutive promoter is routinely used in vector construction in plant genetic engineering.It raises controversy due to the safety issue of transgenic food.The constitutive promoter was replaced by a tissue-specific promoter that aimed to regulate downstream gene expression as well as solve the safety issues of transgenic food.Homology clone method was used to amplify 2 278 bp long Pmsg fragments of pod-specific promoter from cultivated soybean Suinong 14.Such fragments had 99% homologous with the msg promoter of soybean in GenBank.Then both pod-specific expression vector pBMBt and seed-specific expression vector pBGBt were constructed.Within the vector,the modified Cry1Iem gene expression was driven by promoter Pmsg and seed-specific promoter PGy2,respectively.These expression vectors are useful for genetic engineering studies of resist soybean bean pod borer.