Cell‐to‐cell trafficking of cucumber mosaic virus movement protein:green fluorescent protein fusion produced by biolistic gene bombardment in tobacco
Cell‐to‐cell trafficking of cucumber mosaic virus movement protein:green fluorescent protein fusion produced by biolistic gene bombardment in tobacco
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DOI：
10.1046/j.1365-313x.1997.12051223.x
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发表时间：
1997-11
期刊：
影响因子：
7.2
通讯作者：
A. Itaya;Heather Hickman;Y. Bao;R. S. Nelson;B. Ding
中科院分区：
文献类型：
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作者：
A. Itaya;Heather Hickman;Y. Bao;R. S. Nelson;B. Ding
概述以前的显微注射研究表明，一些重组病毒运动蛋白和植物蛋白产生和纯化的大肠杆菌可以在细胞之间的交通。然而，通过微量注射E.这些蛋白质在植物中产生时，大肠杆菌对这些蛋白质的真实的功能的认识受到了质疑。在这项研究中，特定的基因构建体被传递到烟草（Nicotiana tabacum var Samsun）叶表皮的基因枪轰击在植物生产的绿色荧光蛋白（GFP）和黄瓜花叶病毒3a运动蛋白（3a MP）和GFP之间的各种融合。与此相反，3a MP：GFP融合蛋白从大约一半的产生它的细胞移动到邻近的细胞。运动也发生在4°C。突变体3a MP：GFP在所有情况下都不能进行细胞间运动。以这种方式产生的3a MP：GUS融合蛋白也从细胞移动到细胞。我们的数据提供了直接的证据，植物中产生的特定病毒蛋白可以在细胞之间运输。此外，我们的数据表明，CMV 3a MP包含一个信号的运输。我们的方法简单有效，在研究胞间连丝介导的大分子转运方面有许多潜在的应用。
Summary Previous micro-injection studies showed that some recombinant viral movement proteins and plant proteins produced in and purified from Escherichia coli could traffic from cell to cell. However, the relevance of these findings obtained by micro-injecting proteins produced in E. coli to the real functions of these proteins when produced in planta has been questioned. In this study, specific gene constructs were delivered by biolistic bombardment into tobacco (Nicotiana tabacum var Samsun) leaf epidermis for in planta production of the green fluorescent protein (GFP) and various fusions between the cucumber mosaic virus 3a movement protein (3a MP) and GFP. Free GFP remained in cells producing it. In contrast, 3a MP:GFP fusion protein moved from approximately half of the cells producing it into neighboring cells. The movement also occurred at 4°C. A mutant 3a MP:GFP was incapable of cell-to-cell movement in all cases. A 3a MP:GUS fusion protein produced in this manner also moved from cell to cell. Our data provide direct evidence that specific viral proteins produced in planta can be transported between cells. Furthermore, our data suggest that the CMV 3a MP contains a signal for transport. Our approach is simple and efficient and has many potential applications in studying plasmodesma-mediated macromolecular transport.