Figure 4. Yeast two-hybrid screen using mECP as bait and a human brain cDNA library identified CPEc.

mECP was cloned into bait vector pGBKT7. Yeast strain AH109 was co-transformed with mCPE–pGADT7 or CPEc–pGADT7. (A) Transformants were spread on to SD-Ade-His-Leu-Trp medium plates. Vigorous growth was observed for cells co-expressing the mECP bait plasmid and two CPE clones at the high-strength selection plate. 1, mCPE; 2, CPEc. (B) The expression of mECP–BD and CPE–AD fusion proteins in yeast cells was detected by Western blotting with anti-c-myc and anti-HA antibody respectively. mCPE–AD (upper panel, lane 1), CPEc–AD (upper panel, lane 2) and mECP–BD (lower panel, lanes 1 and 2) fusion proteins were expressed in yeast cells.