Construction Of The Eukaryotic Expression Plasmid Of Human FBP17 Gene And Its Expression In LO2 Cells

Objective: To construct the eukaryotic expression plasmid p3×FLAG-CMV-10/FBP17 and transfect p3×FLAG-CMV-10/FBP17 into LO2 cells, and verify high expression of FBP17 of the transfected LO2,as the solid foundation for our further study of the role of FBP17 in the cell signaling pathway and the effect to morphological remodeling.Methods: 1.To construct p3×FLAG-CMV-10/FBP17 recombinant plasmid, restriction enzyme digestion was performed to get the entire fragment of human FBP17 gene, and the gene was inserted into the eukaryotic expression vector p3×FLAG-CMV-10 to form the new construct donated p3×FLAG-CMV-10/FBP17 after purification. 2.To transfect recombinant plasmid p3×FLAG-CMV-10/FBP17 into human LO2 cells by liposome method. 3. To identify high expression of FBP17 in LO2 cells that plasmid p3×FLAG-CMV-10/FBP17 was transfected in. And XTT assay was used to detect the changes in cell multiplication. Results: (1) The eukaryotic expression plasmid p3×FLAG-CMV-10/FBP17 was successfully constructed.(2) The eukaryotic expression plasmid p3×FLAG-CMV-10/FBP17 was successfully transfected into LO2,and the protein of FBP17 in the transfected LO2 was successfully identified.(3) No significant changes in cell multiplication were observed in these transfected cells.