Identification of interacting protein of FOXP3 by yeast two-hybrid system / 第三军医大学学报

ABSTRACT

Objective To screen the proteins interacting with FOXP3 in yeast two-hybrid system. Methods The "bait plasmid" pGBKT7 (named as pGBKT7-FOXP3) was constructed successfully. Using FOXP3 as bait, a human liver cDNA library was screened and the proteins interacting with FOXP3 were searched. The false positive clones were discarded by one to one yeast two-hybrid system, and the positive clones were sequenced and analyzed by bioinformatic methods. Results The bait plasmid pGBKT7-FOXP3 was constructed successfully and there was no self-activation or toxicity in AH109. Three proteins had been found in our system to be able to interact with FOXP3. They were tumor protein D52, splicing factor 3b subunit 1 and one hypothetical protein. Conclusion FOXP3 interacts with tumor protein D52, splicing factor 3b subunit 1 and one hypothetical protein, all of which may interfere in cell metabolism and function of T cell.