Abstract:
Objective To clone the total length cDNA of TFPI22 gene and const ruct it s eukaryotic expres2 sion vector. Then the eukaryotic expression vector was t ransfected into pancreatic carcinoma cell line Panc21, the expression of TFPI22 was detected. Methods The TFPI22 gene was amplified by RT2PCR f rom human placenta tissue and was inserted into eukaryotic expression vector p EGFP2C1, then the eu2 karyotic expression vector p EGFP2C12TFPI22 was const ructed. The recombinant plasmid was then t rans2 fected into pancreatic carcinoma cell line Panc21 cells. The expression of TFPI22 in t ransfected cells was detected by Western blot . Results By the use of RT2PCR, the f ragment of 708 bp of TFPI22 gene was successfully amplified. After being inserted into the vector, enzyme digestion analysis and DNA sequen2 cing showed that the target gene was cloned into recombinant vector successfully. Af ter being t ransfected into Panc21 cell, green fluorescence was emitted f rom t ransfected cells under fluorescent microscope. Western blot analysis revealed that the TFPI22 gene could be expressed stably in the t ransfected cells. Conclusion The eukaryotic expression vector p EGFP2C12TFPI22 was successfully const ructed. The Panc21 cell that could express TFPI22 stably was obtained. It was also confirmed that the TFPI22 gene could be expressed in Panc21 cell with high2performance and stability. These established a foundation for further study of TFPI22 in the invasion and migration of pancreatic carcinoma cells.