Yeast two-hybrid analysis showing interaction between Mep33 and Pir2.
Full-length Mep33 was fused to the LexA DNA-binding domain in pTlexA4 vector (BD-Mep33). The yeast reporter strain L40 cells were co-transformed with BD-Mep33 and various full-length proteins fused to the GAL4 activating domain in pGAD424 vector (AD) as indicated. The transformants were selected on double (-Leu-Trp) dropout media. AD-X represents empty pGAD424 vector as a auto-activation control of BD- Mep33. Control represents co-transformant with AD-X and BD-Rmn1, which is known to interact, as a positive control. (A) Expression of the reporter gene, His3, is monitored by growth on triple dropout media lacking histidine, leucine, and tryptophan (SD-LTH media). (B) Expression of the reporter gene, LacZ, is monitored on double (-Leu-Trp) dropout media containing 5-bromo-4-chloro-3-indolyl-α-D-galactopyranoside (X-gal media).
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