Expression of HLH domain-deleted Id2 protein in yeast cells and detection of its self-activation

Objective:To construct the bait vector of helix-loop-helix（HLH）-deleted Id2 for yeast two-hybrid system and to detect its effect on the growth of yeast cells and the self-activation of reporter genes.Methods:HLH-deleted Id2 was amplified by overlap extension PCR and subsequently cloned into pGBKT7 to construct the bait plasmid for yeast two-hybrid system.These recombinant plasmids were transfected into AH109 yeast cells and their effect on reporter genes and the growth of yeast calls were measured.Results:The fragment encoding HLH-deleted Id2 was successfully amplified and sequenced to confirm its correct expression frame in pGBKT7 plasmid.HLH-deleted Id2 was correctly expressed in yeast cells,both the toxicity and self-activation of which were not observed.Conclusion:The HLH-deleted Id2 interaction proteins can be screened in yeast two-hybrid system.