Figure 1
Yeast surface display of evasins. (A) Arrangement of evasin expression constructs (not to scale). Evasin fusion peptides were either tagged at the N-terminus with a AGA2 peptide (top), or at the C-terminus with SAG1 (middle) or AGA2 peptides (bottom). The position of antibody epitope tags (HA, MYC, FLAG), and the (Gly4Ser)3 linker (G4S3) is also indicated. Evasin fusion expression was driven by a Gal1p promoter. (B) Fluorescence profiles (red curves) of yeast displaying evasin 1 (left), evasin 4 (middle) and evasin 3 (right) panels incubated with indicated biotinylated chemokines and streptavidin-AF647(red curves). Data show a representative experiment of three biological repeats. Surface display tags were placed at the evasin N-terminus (AGA2, top panels) or at the evasin C-terminus (SAG1, bottom panels). Y-axis shows cell count (side scatter), and x-axis the fluorescence intensity on a log-scale. The blue curve is the profile of yeast containing a negative control surface display vector plasmid treated as above, and is used to determine background. The percentages of cells exceeding background levels in evasin displaying yeast are indicated. (C) Fluorescence profiles (red curves) of yeast displaying evasin 4 incubated with biotinylated CCL5 and streptavidin-AF647 (red curves). Data show a representative experiment of three biological repeats. Surface display tags were placed at the evasin N-terminus (AGA2, left panel) or at the evasin C-terminus (SAG1, AGA2, middle and right panels respectively). y-axis shows cell count, and x-axis the fluorescence intensity on a log-scale. The blue curve is the profile of yeast containing a negative control surface display vector plasmid treated as above, and is used to determine background. The percentages of cells exceeding background levels in evasin 4 displaying yeast are indicated.
