Drakakaki G, Christou P, Stöger E
Molecular Biotechnology Unit, John Innes Centre, Norwich, UK.
Transgenic Res. 2000 Dec;9(6):445-52. doi: 10.1023/a:1026534009483.
We used particle bombardment to produce transgenic wheat and rice plants expressing recombinant soybean ferritin, a protein that can store large amounts of iron. The cDNA sequence was isolated from soybean by RT-PCR and expressed using the constitutive maize ubiquitin-1 promoter. The presence of ferritin mRNA and protein was confirmed in the vegetative tissues and seeds of transgenic wheat and rice plants by northern and western blot analysis, respectively. The levels of ferritin mRNA were similar in the vegetative tissues of both species, but ferritin protein levels were higher in rice. Both ferritin mRNA and protein levels were lower in wheat and rice seeds. ICAP spectrometry showed that iron levels increased only in vegetative tissues of transgenic plants, and not in the seeds. These data indicate that recombinant ferritin expression under the control of the maize ubiquitin promoter significantly increases iron levels in vegetative tissues, but that the levels of recombinant ferritin in seeds are not sufficient to increase iron levels significantly over those in the seeds of non-transgenic plants.
我们利用粒子轰击技术培育出了表达重组大豆铁蛋白的转基因小麦和水稻植株，大豆铁蛋白是一种能够储存大量铁的蛋白质。通过逆转录聚合酶链反应（RT-PCR）从大豆中分离出cDNA序列，并使用组成型玉米泛素-1启动子进行表达。分别通过Northern印迹分析和Western印迹分析，在转基因小麦和水稻植株的营养组织和种子中证实了铁蛋白mRNA和蛋白质的存在。两种植物营养组织中铁蛋白mRNA的水平相似，但水稻中铁蛋白的蛋白质水平更高。小麦和水稻种子中铁蛋白mRNA和蛋白质的水平均较低。电感耦合等离子体质谱法（ICAP）分析表明，铁含量仅在转基因植物的营养组织中增加，而在种子中未增加。这些数据表明，在玉米泛素启动子控制下的重组铁蛋白表达显著提高了营养组织中的铁含量，但种子中重组铁蛋白的水平不足以使铁含量比非转基因植物种子中的铁含量显著增加。
