Shah D M, Horsch R B, Klee H J, Kishore G M, Winter J A, Tumer N E, Hironaka C M, Sanders P R, Gasser C S, Aykent S, Siegel N R, Rogers S G, Fraley R T
Science. 1986 Jul 25;233(4762):478-81. doi: 10.1126/science.233.4762.478.
The herbicide glyphosate is a potent inhibitor of the enzyme 5-enolpyruvylshikimate- 3-phosphate (EPSP) synthase in higher plants. A complementary DNA (cDNA) clone encoding EPSP synthase was isolated from a complementary DNA library of a glyphosate-tolerant Petunia hybrida cell line (MP4-G) that overproduces the enzyme. This cell line was shown to overproduce EPSP synthase messenger RNA as a result of a 20-fold amplification of the gene. A chimeric EPSP synthase gene was constructed with the use of the cauliflower mosaic virus 35S promoter to attain high level expression of EPSP synthase and introduced into petunia cells. Transformed petunia cells as well as regenerated transgenic plants were tolerant to glyphosate.
除草剂草甘膦是高等植物中5-烯醇式丙酮酸莽草酸-3-磷酸（EPSP）合酶的有效抑制剂。从过量产生该酶的耐草甘膦矮牵牛杂交细胞系（MP4-G）的互补DNA文库中分离出一个编码EPSP合酶的互补DNA（cDNA）克隆。由于该基因20倍的扩增，该细胞系过量产生EPSP合酶信使核糖核酸。利用花椰菜花叶病毒35S启动子构建了一个嵌合EPSP合酶基因，以实现EPSP合酶的高水平表达，并将其导入矮牵牛细胞。转化的矮牵牛细胞以及再生的转基因植物对草甘膦具有耐受性。
