Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey from embryo cDNA expression library).
Two-hybrid system: yeast Stratagene CytoTrap
Interaction of prey with bait targets hSos domain to membrane, activating Ras and permitting mutant cdc25H yeast to grow at 37oC.
The majority of yki protein is cytoplasmic. To avoid isolating these cytoplasmic factors and instead identify proteins involved in yki transcriptional functions, yki was immunoprecipitated from nuclear extracts.
Source was nuclear extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in bacterial system.
Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.