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. 2016 Oct 20;6(12):4115–4125. doi: 10.1534/g3.116.033035

Figure 5.

Figure 5

Expression of Venus-3FLAG-tagged peptides and proteins from one-promoter constructs. Strain CC-124 was transformed with the constructs from the indicated plasmids. (A) Analysis of protein expression by SDS-PAGE and Western blotting using an anti-FLAG antibody as described in Figure 2D. In all cases but pMO424, one transformant for each construct was chosen at random for analysis. For pMO424, 192 transformants were screened by fluorescence microscopy, and the one with the strongest signal was picked for further analysis. Predicted molecular weights of the fusion proteins are shown. *, the 1.4-kDa difference in predicted molecular weight of Lifeact-Venus-3FLAG expressed from the pMO424 and pMO459 constructs is due to the seven additional amino acids (MARRFEV) in the latter encoded by sequences added during the construction. (B, C) Localization of Venus-3FLAG-tagged VFL2 [(B); the same transformant as in (A)] and PMH1 [(C); transformation with the construct from pMO515]. Cells were observed using the spinning-disk microscope. Bars, 5 µm.