Maiti I B, Hunt A G, Wagner G J
Department of Agronomy, University of Kentucky, Lexington 40546-0091.
Biochem Biophys Res Commun. 1988 Jan 29;150(2):640-7. doi: 10.1016/0006-291x(88)90440-8.
A binary plasmid was constructed to contain the mouse metallothionein c-DNA, the constitutive 35S promoter from cauliflower mosaic virus, the polyadenylation signal from the pea rbcS-E9 gene and several selectable markers. The plasmid was transferred to Agrobacterium tumefaciens and the leaf disc method was used to transform tobacco. Callus and shoots were regenerated in the presence of kanamycin and transformed plants were obtained. Southern, Northern and Western blot analysis demonstrated integration and expression of the metallothionein gene in transformed callus and transgenic plants. The gene is transmitted to and expressed in seed derived progeny as a dominant Mendelian trait.
构建了一种二元质粒,其包含小鼠金属硫蛋白cDNA、来自花椰菜花叶病毒的组成型35S启动子、来自豌豆rbcS-E9基因的聚腺苷酸化信号以及几个选择标记。将该质粒转移到根癌农杆菌中,并使用叶盘法转化烟草。在卡那霉素存在的情况下再生出愈伤组织和芽,并获得了转化植株。Southern、Northern和Western印迹分析表明金属硫蛋白基因在转化的愈伤组织和转基因植株中整合并表达。该基因作为显性孟德尔性状传递给种子衍生的后代并在其中表达。