大豆Gy1基因种子特异性表达载体的构建及在玉米中的转化

实验构建了2个大豆贮藏蛋白Gy1基因种子特异性表达载体,并将其分别转入玉米愈伤组织中。载体1以潮霉素为筛选标记,将Gy1基因片段插入到含有种子特异启动子的植物表达载体pCAMBIA1301-7αP上,得到种子特异性表达载体pCAMBIA1301-7αP-Gy1;载体2以BADH为筛选标记,将Gy1基因插入到带有耐盐基因BADH的植物表达载体pCAMBIA1301-BADH上,得到安全性无抗生素选择标记的种子特异性表达载体pCAMBI-A1301-sbeⅡb-Gy1。通过农杆菌介导法将其分别导入玉米自交系H99的愈伤组织中,共获得132株再生植株;经过PCR检测,初步证明获得了34株转基因阳性植株。

Construction of the Seed-specific Expression Vector Harboring Gy1 Gene From Soybean and Transformation of Corn

Two seed-specificity expression vectors harboring seed storage protein Gy1 gene of soybean were con-structed and the two vectors were transformed into embryogenic calluses of corn.Vector1, pCAMBIA1301-7αP-Gy1, was constructed by inserting Gy1 gene into pCAMBIA1301-7αP with selectable marker hyg, and vector 2, pCAMBI-A1301-sbe Ⅱ b-Gy1, was constructed by inserting Gy1 gene into pCAMBIA1301-BADH with salt-tolerance se-lectable marker BA DH.The two vectors were transformed into embryogenic calluses of maize "H99" mediated with a-grobacterium.Thirty-four transgenic plants were confirmed by PCR analysis from 132 putative transgenic plants.