Two-hybrid system: yeast GAL4-BD/GAL4-AD

siz-RA and siz-RB differ in the first 116 aa but retain conserved domains including the IQ motif. Since the IQ motif is not required for CadN binding, the authors reason that the shared 207 aa between siz-RA and siz-RB are essential for binding to CadN.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey from embryo cDNA expression library).

Transgenic flies producing each of the siz and CadN fusion constructs were created such that the construct were expressed under the control of a Mef2 driver. Interaction between siz and CadN reconstituted the YFP protein which was then visualized using anti-GFP antibody. Flies expressing the siz or CadN constructs alone did not show signal when stained with anti-GFP.

Interaction in vivo; source was embryos of transgenic fly line; proteins produced from transgenic constructs.