Subcellular distribution of mutant movement proteins of Cucumber mosaic virus fused to green fluorescent proteins, by T. Canto and P. Palukaitis
Journal of General Virology vol. 86, part 4, pp. 1223 – 1228
Supplementary Figure. Localization of MP–red fluorescent protein and tubulin–GFP in epidermal cells. Genes encoding the wild-type (WT) 3a protein, as well as the MPs of the dysfunctional mutant M4 and the conditionally functional mutant M8, were cloned in the binary vector pROK2 and fused at their C termini to the monomeric red fluorescent protein (mRFP; Campbell et al., 2002). These binary constructs were agroinfiltrated into either transgenic Nicotiana benthamiana plants expressing α-tubulin fused to GFP (tua–GFP) (line CB13; Gillespie et al., 2002) for transient expression of the tagged MPs (WT and M4), or non-transgenic N. benthamiana co-infiltrated with a binary construct expressing tua–GFP (Ueda et al., 1999) (M8). At 3 days post-infiltration, the three MP–mRFP fusions showed the same intracellular distribution of fluorescence as when tagged with GFP: discrete spots at the cell wall for WT 3a–mRFP (upper panels) or cytoplasmic and nuclear distribution for both M4–mRFP and M8–mRFP, the former showing a more granular and threadbare appearance than the latter (middle and lower panels, respectively). In neither case did the red fluorescence appear to target the microtubule filaments highlighted green by tua–GFP. Left panels show whole cells; right panels show enhanced detail of the intracellular microtubular net. Bars, 50 µm.
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