Western blot analysis of expression of Fap1 and Gap3 variants. Gap3 deletion mutants, Δ62–67 mutant (Lane 1), Δ144–157 mutant (Lane 2), wild type strain (Lane 3), gap3 allelic replacement mutant VT1619 (Lane 4), gap3 complemented strain VT1732 (Lane 5) and a control strain (VT1619 transformed with empty vector pVT1666) (Lane 6) were subjected to Western blot analysis with the use of MAb D10 (A). Expression of Fap1 by Gap3 site-directed mutants, V33R, F35H, N37I, P38R, S42L, N54I, R59L, P62G, D63V, L64R, P65R, I66N, L67T and L75S (Lanes 1–14) and wild type strain (Lane 15) probed with MAb D10 (B) and MAb F51 (C). Expression of Gap3-GFP by gap3 site-directed mutants, Gap3 L64R (Lane 1), P65R (Lane 2), I66N (Lane3), L67T (Lane4), Gap3 complemented strain VT1732 (Lane 5), control (VT1619 with empty vector pVT1666) (Lane 6) and gap3 mutant VT1619 (Lane 7) probed with anti-GFP MAb (C). Gap3-GFP fusion proteins migrate at 48 kDa; GFP protein migrates at 27 kDa.