The Screen Of Human Trefoil Factor 1 Interacting Protein Gene

Objective To investigate the expression of human trefoil factor 1 (hTFF1) in normal and gastric carcinoma mucosa and screen the interacting protein of hTFF1 in human gastric carcinoma cells. Methods The expression of hTFF1 in normal and gastric carcinoma mucosa was assessed by immunohistochemical method. The hTFF1 gene was obtained from normal gastric mucosa and pDEST32-hTFF1, the Yeast Two Hybrid bait vector of hTFF1 was constructed. The pDEST22-gastric carcinoma cDNA libray vectors, the prey vectors of the Yeast Two Hybrid system were constructed by BP reaction and LR reaction at the same time. Then pDEST32-hTFF1 and pDEST22-gastric carcinoma cDNA libray vectors were transfected into MaV203 yeast cells and screened by defected solid nutrient medium.The result was analyzed by nucleotide data base and bioinformatics technique. To validate the interaction between the candidate proteins and hTFF1 protein, the bait plasmids expressing candidate proteins were constructed. The prey plasmids for Reverse Yeast Two Hybrid system expressing hTFF1 protein was constructed as well. The reconstituted bait and prey plasmids were cotransform into the yeast cell MaV203 to testify their interaction.Results The expression of hTFF1 in gastric carcinoma was obviously lower than that in normal gastric mucosa. The hTFF1 gene was successfully obtained from normal human gastric mucosa. The pDEST32-hTFF1 vector and the pDEST22-gastric carcinoma cDNA libray vectors were successfully constructed. 24 positive clones were obtained from the screen, and 1 was successfully sequenced. The FAM38A protein(Homo sapiens family with sequence similarity 38, member A) was obtained. The interaction between FAM38A and hTFF1 was certificated by reverse yeast two-hybrid.Conclusion hTFF1 is likely to restrain the generation and progression of gastric cancer. Successfully screen and clone FAM38A,the TFF1 interacting protein genes from the gastric carcinoma cells which were very important for further investigation about the function of TFF1 in the genesis and progress of gastric carcinoma.