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. 2009 Feb 1;23(3):331–344. doi: 10.1101/gad.507609

Figure 4.

Figure 4.

Translation of the second frame in pYF constructs in yeast depends on translation of the first frame. (A) Schematic drawing of the upstream frame of plasmid pYF72 compared with pYF21A (not drawn to scale). The TURBS (T) is highlighted in gray and the two essential motifs are indicated. Insertion of four extra nucleotides (written in bold) at a BglII site (BglII*) 275 nt upstream of the ORF2 stop codon results in a frameshift and premature termination of ORF2 translation. The gray bars below indicate the proteins encoded by ORF2. The VP1* expressed from the frameshift mutant pCH72 is truncated by 70 amino acids, and the C-terminal 18 amino acids of the protein represent non-FCV residues, resulting from the introduced frameshift (light-gray area). Note that ORF3 is not shown in the scheme. (B) The proteins translated in the yeast system are shown in a Western blot analysis with antisera against VP1 (top panel) or GFP (bottom panel). (Gal-Pro) Yeast galactose-promoter.