糖皮质激素诱导绿色荧光蛋白在烟草中的表达  

Glucocorticoid-Inducible Green Fluorescent Protein Expression in Tobacco Plants

作　　者：王俊杰[1] 宋晔华[1] 范亚军[2] 王艳[1] 孙少广[1] 许多[1] 朱筱娟[1] 王兴智[1] 

机构地区：[1]东北师范大学遗传与细胞研究所,长春130024 [2]长春师范学院生命科学学院,长春130032

出　　处：《吉林农业大学学报》2008年第4期427-435,共9页Journal of Jilin Agricultural University

基　　金：国家植物转基因中试与产业化基地(吉林)(JY03-B-17)

摘　　要：糖皮质激素诱导体系具有操作简单、化学物质稳定、可控性好等显著优点,被认为是最有应用潜力的化学诱导体系。利用农杆菌介导方法,将携带诱导表达元件的双元载体pIndex3-NTR-mgfp5转入野生型烟草,转录因子GVG的基因由35S组成型启动子控制表达,绿色荧光蛋白基因mgfp5作为诱导表达的目的基因在诱导启动子的控制下表达。结果表明:绿色荧光蛋白的转录受到诱导剂的严格调控。同时发现,浸根诱导法对基因表达的调控首先表现在新生叶片上,在老叶片中则相对滞后。此外,在质粒构建过程中,在mgfp5基因两端分别加上马铃薯X病毒(PVX)的非翻译区序列5NTR和3NTR,结果非翻译区序列并未抑制绿色荧光蛋白的表达。在建立的糖皮质激素诱导表达体系中,GVG系统对诱导剂具有很高的专一性,并且对基因表达的控制非常严格,几乎没有本底表达,可以从时间、空间及数量上对目的基因进行精确调控。这一体系的建立为今后利用化学诱导的方法进行理论和应用研究奠定了基础。The glucocorticoid-mediated induction system is considered to be the most potential chemical induced system in transgenic application, which has some prominent advantages, including easy manipulation, stable inducer transcription factor GVG gene under the control of 35S promoter and the mgfp5 transcribed from an inducible promoter and convenient to control. In our study, the binary vector pIn-dex-NTR-mgfp5, carrying the glueocorticoid inducible expression elements, was transferred into A- grobacterium and wild-type tobacco plants. In this system, the transcription factor GVG gene under the control of 35S promoter and the mgfp5 transcribed from an inducible promoter were used as the target gene. The results indicated that the mgfp5 expression was rigorously controlled, and the young leaves were more sensitive to the inducer than the old ones. The non-translated regions （5NTR and 3NTR ） of the PVX was inserted into the flanked mgfp5 gene and they did not suppress the mgfp5 expression. After detection, it was comfirmed that the glucocorticoid inducible expression system was established. The GVG system was of high stringency and specificity, and could control the transgenic expression temporally, spatially and quantitatively induced by exogenous chemicals. The establishment of this system may offer the possibility for both theoretical and practical applications by the chemical inducible system.

关 键 词：糖皮质激素诱导系统 地塞米松 绿色荧光蛋白 非翻译区 

分 类 号：Q786[生物学—分子生物学] S572[农业科学—烟草工业]