Phytogen, 101 Waverly Drive, Pasadena, California 91105.
Plant Physiol. 1992 Feb;98(2):458-64. doi: 10.1104/pp.98.2.458.
The expression of maize (Zea mays) phophoenolpyruvate carboxylase (PPC) gene constructions was studied in transgenic tobacco plants (Nicotiana tabacum). Where transcription was under the control of a maize PPC gene promoter, a low level of aberrantly large PPC transcript was detected. Analysis of this PPC transcript indicated that transcription initiation occurs upstream of the normal site. Despite the aberrant transcription initiation, expression of the PPC transcript was still light-regulated. Higher levels of maize PPC transcript of the correct size were obtained with a chimeric gene construction containing a tobacco (Nicotiana plumbaginifolia) chlorophyll a/b binding protein gene promoter. The PPC activities in the leaves of these transgenic plants were up to twofold higher than those of nontransformed plants. Two forms of PPC with different kinetic properties were identified in leaf extracts of the transgenic plants: one form with a high apparent K(m) for phosphoenolpyruvate (maize isozyme), and a second form exhibiting a low apparent K(m) (tobacco isozyme). Biochemical analyses of these plants indicated that the transgenic plants had significantly elevated levels of titratable acidity and malic acid. These biochemical differences did not produce any significant physiological changes with respect to photosynthetic rate or CO(2) compensation point.
在转基因烟草植物(Nicotiana tabacum)中研究了玉米(Zea mays)磷酸烯醇丙酮酸羧化酶(PPC)基因构建体的表达。在玉米 PPC 基因启动子的控制下,检测到低水平的异常大的 PPC 转录本。对该 PPC 转录本的分析表明,转录起始发生在正常位点的上游。尽管转录起始异常,但 PPC 转录本的表达仍受光调控。使用包含烟草(Nicotiana plumbaginifolia)叶绿素 a/b 结合蛋白基因启动子的嵌合基因构建体获得了更高水平的正确大小的玉米 PPC 转录本。这些转基因植物叶片中的 PPC 活性比非转化植物高两倍。在转基因植物的叶片提取物中鉴定出两种具有不同动力学特性的 PPC 形式:一种形式对磷酸烯醇丙酮酸(玉米同工酶)具有高表观 Km,另一种形式表现出低表观 Km(烟草同工酶)。对这些植物的生化分析表明,转基因植物的可滴定酸度和苹果酸水平显著升高。这些生化差异在光合速率或 CO2 补偿点方面没有产生任何显著的生理变化。