Virus-mediated delivery of the green fluorescent protein to the endoplasmic reticulum of plant cells
Virus-mediated delivery of the green fluorescent protein to the endoplasmic reticulum of plant cells
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DOI：
10.1046/j.1365-313x.1996.10050935.x
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发表时间：
1996-11-01
期刊：
影响因子：
7.2
通讯作者：
Oparka, KJ
中科院分区：
文献类型：
--
作者：
Boevink, P;SantaCruz, S;Oparka, KJ
利用基于病毒的表达系统将绿色荧光蛋白(GFP)定向到活植物细胞的内质网(ER)。将来自储存蛋白patatin的信号肽与GFP的氨基末端融合，将ER保留信号KDEL与羧基末端融合。将嵌合的GFP基因插入马铃薯病毒X基因表达载体和体外转录本，构建了重组病毒基因组。接种于本氏烟草和四裂叶烟上。在感染病毒的细胞中，GFP既针对皮质ER，也针对在细胞质中流动的ER元件的移动系统。此外，大量含有GFP的包裹体也很明显。这些包涵体是可移动的，但仍与内质网中的元素密切相关。用膜电位敏感染料对内质网进行染色，证实了GFP是针对内质网的，并讨论了病毒介导的递送系统在植物内膜系统研究中的应用。
The green fluorescent protein (GFP) was targeted to the endoplasmic reticulum (ER) of living plant cells using a virus-based expression system. The signal peptide from the storage protein, patatin was fused to the amino-terminus of the GFP while the ER retention signal KDEL was fused to the carboxy-terminus. The chimaeric gfp cDNA was inserted into a potato virus X-based expression vector and in-vitro transcripts, representing the recombinant viral genome. were inoculated on to Nicotiana benthamiana and IV. clevelandii plants. In virus-infected cells, the GFP was targeted both to the cortical ER and to a mobile system of ER elements which underwent streaming in the cytoplasm. In addition, a population of GFP-containing inclusions was apparent. These inclusions were motile but remained closely associated with elements of the ER. Staining of the ER with membrane potential-sensitive dyes confirmed that the GFP had been targeted to the ER, The utility of virus-mediated delivery systems in studies of the plant endomembrane system is discussed.