The Screen Of Human Trefoil Factor 2 Interacting Protein Gene In CDNA Library Of Gastric Cancer Cell

Objective:Screen ing and cloning of human trefoil factor 2(trefoil factor family 2, TFF2) interacting protein gene in cDNA library of gastric cancer cell.Methods:Verify TFF2mRNA expression in gastric cancer by RT-PCR method;First amplify TFF2 gene by PCR,second clone TFF2 into yeast expression vector pDEST32 after Not I and Sal I double digested to construct the TFF2 bait plasmid finally;confirm the fusion protein expression of TFF2 bait plasmids in yeast cells by western blot; transfect The TFF2 bait plasmid and prey plasmid of human gastric cancer-cDNA library into Mav203 yeast cells.The positive clones were screened through auxotrophic medium (Ura,His) and X-gal,then extracted for sequencing,and for analysising by the protein databases and bioinformatics technology.Results:TFF2 mRNA was successfully expressed in the gastric cancer;pDEST32-TFF2 bait plasmid was successfully constructed;The TFF2 baitplasmid precisely expressing the TFF2-6AL4DBD fusion protein was confirmed by western blot method;35 positive clones were obtained through the yeast two-hybrid technology screening in the gastric cDNA library,of which 16 were successfully sequenced,including 12 known and 4 unknown functional genes.Conclusion:TFF2 possibly play an important role in the occurrence and development of gastric cancer;A variety of TFF2 interacting protein gene were screened from the gastric cancer CDNA library;Further analysising the function of interacting proteins may contribute to excavate the possibly mechanisms of TFF2' s fuction in the occurrence and development of gastric cancer.