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. 2003 Mar;23(5):1750–1763. doi: 10.1128/MCB.23.5.1750-1763.2003

FIG. 4.

FIG. 4.

Two-hybrid interactions among Far3 and Far7 to Far9. Two-hybrid tests were performed as described in Materials and Methods. Fusion proteins were tested in pairwise combinations consisting of a bait (Gal4BD fused to the N terminus of full-length Far3 or Far7) and a target (Gal4AD alone or Gal4AD fused to the N terminus of a Far protein or protein fragment). BD-Far3 and BD-Far7 baits were expressed from pSL2785 and pSL2786, respectively. Target proteins were expressed from plasmids as follows: AD alone from pGAD-C1; AD-Far7A (AD plus the C terminus of Far7, amino acids 65 to 222) from pSL2789; AD-Far8 (AD fused to full-length Far8) from pSL2790; AD-Far9B (AD plus the internal fragment of Far9, amino acids 138 to 572) from pSL2792. Interactions were assayed by patching independent isolates in duplicate to SD-Trp Leu and then replica plating to SD-Trp Leu His and SD-Trp Leu Ade. Growth on both tester media indicated a positive result. Patch tests were repeated at least four times using different isolates each time; the results of a typical patch test are shown.