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

作     者：Wan Bingbing Shi Yan Huo Keke 

作者机构：State Key Laboratory of Genetic EngineeringInstitute of GeneticsSchool of Life SciencesFudan UniversityShanghai 200433China 

出 版 物：《Frontiers in Biology》 (生物学前沿（英文版）)

年 卷 期：2006年第1卷第2期

页      面：120-126页

核心收录：

学科分类：08[工学] 0812[工学-计算机科学与技术（可授工学、理学学位）] 

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) CNHLPP (2004BA711A19) Shanghai Science and Technology Innovation Program 

主　　题：yeast two-hybrid system protein-protein interaction human liver cDNA library FOXA3 

摘      要：Yeast two-hybrid approach is popularly used nowadays as an important technical method in the field of studying protein-protein *** 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 *** 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 ***,it usually results in failures because it contains the activate domain and can self-activate the reporter *** 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 *** we used this two-hybrid library to screen FOXA3,a hepatocyte nuclear factor,and found out an interacting protein:complement component C3.