Elliott R. C., Dickey L. F., White M. J., Thompson W. F.
Department of Botany and Department of Genetics, North Carolina State University, Raleigh, North Carolina 27695.
Plant Cell. 1989 Jul;1(7):691-698. doi: 10.1105/tpc.1.7.691.
An intact pea gene encoding ferredoxin I (Fed-1) and several chimeric constructs containing portions of Fed-1 were introduced into tobacco plants by Agrobacterium-mediated transformation. The intact gene was correctly transcribed and translated to produce a protein that was imported into the chloroplast and processed to its mature size. Fed-1 mRNA accumulation in these plants was strongly light-dependent, as it is in pea leaves. In chimeric constructs, the Fed-1 promoter was active but no light responses were seen, even when as much as 2 kilobases of 5[prime] -flanking sequence were included. We also failed to observe clear light responses with a construct containing 3[prime] -flanking sequences from Fed-1 attached to a [beta]-glucuronidase gene driven by the cauliflower mosaic virus 35S promoter. However, the transcribed portion of Fed-1 conveyed normal light responsiveness when driven by the 35S promoter. The results are discussed in terms of the hypothesis that light determines Fed-1 mRNA abundance by affecting RNA stability rather than by affecting transcription.
通过农杆菌介导的转化,将一个完整的编码铁氧化还原蛋白I(Fed-1)的豌豆基因以及几个包含Fed-1部分片段的嵌合构建体导入烟草植株。完整基因被正确转录和翻译,产生一种被导入叶绿体并加工成成熟大小的蛋白质。这些植株中Fed-1 mRNA的积累强烈依赖光照,就像在豌豆叶片中一样。在嵌合构建体中,Fed-1启动子具有活性,但未观察到光反应,即使包含多达2千碱基的5′侧翼序列也是如此。我们用一个构建体也未能观察到明显的光反应,该构建体包含来自Fed-1的3′侧翼序列,连接到由花椰菜花叶病毒35S启动子驱动的β-葡萄糖醛酸酶基因上。然而,当由35S启动子驱动时,Fed-1的转录部分表现出正常的光反应性。根据光照通过影响RNA稳定性而非转录来决定Fed-1 mRNA丰度这一假设对结果进行了讨论。