Lee Lan-Ying, Kononov Maria E, Bassuner Burgund, Frame Bronwyn R, Wang Kan, Gelvin Stanton B
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392, USA.
Plant Physiol. 2007 Dec;145(4):1294-300. doi: 10.1104/pp.107.106633. Epub 2007 Oct 11.
We developed novel plasmids and T-DNA binary vectors that incorporate a modified and more useful form of the superpromoter. The superpromoter consists of a trimer of the octopine synthase transcriptional activating element affixed to the mannopine synthase2' (mas2') transcriptional activating element plus minimal promoter. We tested a superpromoter-beta-glucuronidaseA fusion gene in stably transformed tobacco (Nicotiana tabacum) and maize (Zea mays) plants and in transiently transformed maize Black Mexican Sweet protoplasts. In both tobacco and maize, superpromoter activity was much greater in roots than in leaves. In tobacco, superpromoter activity was greater in mature leaves than in young leaves, whereas in maize activity differed little among the tested aerial portions of the plant. When compared with other commonly used promoters (cauliflower mosaic virus 35S, mas2', and maize ubiquitin), superpromoter activity was approximately equivalent to those of the other promoters in both maize Black Mexican Sweet suspension cells and in stably transformed maize plants. The addition of a maize ubiquitin intron downstream of the superpromoter did not enhance activity in stably transformed maize.
我们构建了新型质粒和T-DNA双元载体,其中包含一种经过修饰且更具实用性的超级启动子形式。该超级启动子由附着于甘露碱合成酶2'(mas2')转录激活元件上的章鱼碱合成酶转录激活元件三聚体加上最小启动子组成。我们在稳定转化的烟草(烟草属)和玉米(玉米属)植株以及瞬时转化的玉米黑墨西哥甜玉米原生质体中测试了一个超级启动子-β-葡萄糖醛酸酶A融合基因。在烟草和玉米中,超级启动子在根中的活性均远高于叶。在烟草中,超级启动子在成熟叶中的活性高于幼叶,而在玉米中,该启动子在植株测试的地上部分之间活性差异不大。与其他常用启动子(花椰菜花叶病毒35S、mas2'和玉米泛素)相比,超级启动子在玉米黑墨西哥甜玉米悬浮细胞和稳定转化的玉米植株中的活性与其他启动子大致相当。在超级启动子下游添加玉米泛素内含子并未增强其在稳定转化玉米中的活性。