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 inter...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.展开更多
基金supported by Chinese National key Grant of Basic Research (2001 CB510203)Hi-Tech Research and Development Program of China (2002AA,227011 and 2002BA711A11)+1 种基金CNHLPP (2004BA711A19)Shanghai Science and Technology Innovation Program.
文摘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.
