Figure 1.

Outline of the cloning and mbSUS screen. Selected ORFs were amplified from Arabidopsis seedling cDNA and cloned into pDONR221 (or pDONR221-f1) by Gateway recombination. ORFs that passed the quality control criteria were mobilized into Cub and NubG destination vectors and introduced into THY.AP4 and THY.AP5 yeast strains. Yeast expressing the Cub clones were pre-screened to remove “false positives” and “false negatives.” Remaining Cub clones were mated with NubG clones that had been arrayed into two 1536-well plates. Successful mating was selected on DS media. After 3 days, diploids were replica-plated to IS media selecting for activation of the His3 and Ade2 markers and growth was scored.