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Author:
Date:
2014-10-20
[Abstract] Here, we present a simple and rapid protocol to measure the extent of cell-to-cell movement of RNA viruses in planta. To do that, the green fluorescent protein (GFP) gene was incorporated into the genome of Melon necrotic spot virus (MNSV) as a coat protein (CP) fusion protein using the Thosea asigna virus 2A catalytic peptide (TaV 2a) (Serra-Soriano et al., 2014). TaV 2a allows the co-translational cleavage of the fusion protein resulting in the independent expression of both proteins (Kim et al., 2011). Viral infection was initiated by agro-infiltration of Cucumis melo leaves. At 6-7 days post-infiltration, fluorescent infection foci images were taken with a fluorescent stereo microscope and infection areas were measured using FIJI ...
[摘要] 在这里，我们提出一个简单和快速的协议，以测量RNA病毒在植物中的细胞到细胞运动的程度。 为此，将绿色荧光蛋白（GFP）基因作为外壳蛋白（CP）融合蛋白掺入到Melon坏死斑病毒（MNSV）的基因组中，使用Sucha asigna病毒/Ta> 2A催化肽（TaV 2a）（Serra-Soriano等人，2014）。 TaV 2a允许融合蛋白的共翻译切割，导致两种蛋白的独立表达（Kim等人，2011）。 病毒感染通过对黄瓜叶的农杆菌浸润来启动。 在浸润后6-7天，用荧光立体显微镜拍摄荧光感染灶图像，并使用FIJI软件测量感染面积。
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