Leffel S M, Mabon S A, Stewart C N
University of North Carolina, Greensboro, USA.
Biotechniques. 1997 Nov;23(5):912-8. doi: 10.2144/97235bi01.
Green fluorescent protein (GFP) is increasingly being used in plant biology from the cellular level to whole plant level. At the cellular level, GFP is being used as an in vivo reporter to assess frequency of transient and stable transformation. GFP has also proven to be an invaluable tool in monitoring trafficking and subcellular localization of protein. At the organ level and up, many exciting applications are rapidly emerging. The development of brighter GFP mutants with more robust folding properties has enabled better macroscopic visualization of GFP in whole leaves and plants. One interesting example has been the use of GFP to monitor virus movement in and among whole plants. GFP is also emerging as a powerful tool to monitor transgene movement and transgenic plants in the field. In a proof-of-concept study, tobacco was transformed with a modified version of the GFP gene controlled by a constitutive (35S) promoter. GFP expression in progeny plants ranged from 0% to 0.5%, and approximately 0.1% GFP was the minimal amount needed for unambiguous macroscopic detection. GFP is the first truly in vivo reporter system useful in whole plants, and we project its usefulness will increase even further as better forms of GFP genes become available.
绿色荧光蛋白(GFP)在植物生物学领域的应用正日益广泛,涵盖了从细胞水平到整株植物水平。在细胞水平上,GFP被用作体内报告基因,以评估瞬时转化和稳定转化的频率。GFP还被证明是监测蛋白质运输和亚细胞定位的宝贵工具。在器官及更高层面,许多令人兴奋的应用正迅速涌现。具有更强折叠特性的更亮GFP突变体的开发,使得在整片叶子和整株植物中对GFP进行更好的宏观可视化成为可能。一个有趣的例子是利用GFP监测病毒在整株植物内部及之间的移动。GFP也正成为监测田间转基因移动和转基因植物的强大工具。在一项概念验证研究中,用一个由组成型(35S)启动子控制的GFP基因修饰版本转化烟草。子代植物中GFP的表达范围为0%至0.5%,而约0.1%的GFP是进行明确宏观检测所需的最小量。GFP是首个真正可用于整株植物的体内报告系统,并且我们预计随着更好形式的GFP基因问世,其用途将进一步增加。