Fig. 3. Interactions of mutant arf1p with Arf GAPs in the DupLEX-ATM two-hybrid system. The interactions between N-terminally truncated (N17) Arf1p in the bait vector (pGlida) and Glo3p or Gcs1p in the prey vector (pJG4-5) were tested in the host strain EGY48. The Q71L mutation had been introduced in wild-type (ARF1), arf1-16 mutant (arf1-16) and arf1-17 mutant (arf1-17) genes. Transformants were plated on MVD (-Trp, -His), grown for 2 days, replicated on MVGal (minimal galactose medium) (-Trp, -His, -Leu), MVD (-Trp, -His), and MVD (-Trp, -His, -Leu) plates, and then further incubated at 30°C for 6 days. Empty prey vector (pJG4-5) was used as a negative control.