Adoption of the 2A Ribosomal Skip Principle to Tobacco Mosaic Virus for Peptide Display.
Adoption of the 2A Ribosomal Skip Principle to Tobacco Mosaic Virus for Peptide Display.
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DOI：
10.3389/fpls.2017.01125
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发表时间：
2017
影响因子：
5.6
通讯作者：
Commandeur U
中科院分区：
文献类型：
--
作者：
Röder J;Fischer R;Commandeur U
植物病毒适合作为纳米材料和纳米颗粒的结构单元，因为它们易于修饰，并且可以使用植物或异源表达系统表达和纯化。植物病毒纳米颗粒已被用于疫苗中的表位呈递，用于药物递送，作为纳米球和纳米线，以及用于生物医学成像应用。荧光蛋白融合体已被用于标记植物病毒颗粒。单体非氧依赖性荧光蛋白iLOV可用作绿色荧光蛋白的替代物。在这项研究中，iLOV序列直接或通过甘氨酸-丝氨酸接头与烟草花叶病毒（TMV）外壳蛋白（CP）的C-末端遗传融合，并携带N-末端口蹄疫病毒（FMDV）2A序列。用重组病毒载体接种烟草（Nicotiana benthamiana）植株，实现了系统感染。通过Western印迹分析和透射电子显微镜证实iLOV融合蛋白和杂交颗粒的存在。我们的数据表明，当与FMDV 2A序列组合时，基于TMV的载体适合于产生至少与iLOV一样大的蛋白质。这种方法允许同时产生融合到CP以及游离CP亚基的外源蛋白。
Plant viruses are suitable as building blocks for nanomaterials and nanoparticles because they are easy to modify and can be expressed and purified using plants or heterologous expression systems. Plant virus nanoparticles have been utilized for epitope presentation in vaccines, for drug delivery, as nanospheres and nanowires, and for biomedical imaging applications. Fluorescent protein fusions have been instrumental for the tagging of plant virus particles. The monomeric non-oxygen-dependent fluorescent protein iLOV can be used as an alternative to green fluorescent protein. In this study, the iLOV sequence was genetically fused either directly or via a glycine-serine linker to the C-terminus of the Tobacco mosaic virus (TMV) coat protein (CP) and also carried an N-terminal Foot-and-mouth disease virus (FMDV) 2A sequence. Nicotiana benthamiana plants were inoculated with recombinant viral vectors and a systemic infection was achieved. The presence of iLOV fusion proteins and hybrid particles was confirmed by western blot analysis and transmission electron microscopy. Our data suggest that TMV-based vectors are suitable for the production of proteins at least as large as iLOV when combined with the FMDV 2A sequence. This approach allowed the simultaneous production of foreign proteins fused to the CP as well as free CP subunits.