An improved yeast two-hybrid approach for detection of interacting proteins

Abstract

Yeast two-hybrid approach is popularly used nowadays as an important technical method in the field of studying protein-protein interactions. Although yeast two-hybrid system is obviously advantageous in searching interacting proteins and setting up the network of proteins interaction, not all of proteins can use routine yeast two-hybrid method to search interacting proteins. Many important proteins, such as some nucleoprotein transcriptional factor, carry out the regular method and construct the bait-BD vector to screen the library containing AD vector. However, it usually results in failures because it contains the activate domain and can self-activate the reporter gene. In this study, we changed the research strategy, fused the bait gene (FOXA3) with the AD vector to screen the library containing BD vector, so that we constructed a two-hybrid library containing BD vector and can bypass the interference of self-activation. And we used this two-hybrid library to screen FOXA3, a hepatocyte nuclear factor, and found out an interacting protein: complement component C3.