Abstract:
Objective: To construct a green fluorescence protein (GFP) expression vector combined with the CX -CL12-KDEL gene and study its expression in human tongue cells. Methods:The combined genes CXCL12-KDEL were am-plified by PCR from human genomic DNA. After purification it was ligated into the 19T vector then inserted into pIRES 2-EGFP by homologous recombination to construct the CXCL12-KDEL-pIRES2-EGFP vector. The constructed plas-mids were identified by BglⅡ/Sal Ⅰrestriction digestion and sequence analysis. Recombinant plasmids were transfected into the Tb 3.1 cell line by Lipofectamine (TM 2000). Cells were observed at 48and 72hours after transfection using an in -verted fluorescence microscope. The expression of CXCL 12was identified by Western blot. Results: The correct construct-ed plasmids were confirmed by restriction digestion and sequence analysis. The results of cell transfection indicated that the GFP could be observed in Tb 3.1 cells. The Tb 3.1 cells can express E-tag protein when transfected with the constructed plasmid as shown by Western blot. Conclusion:The GFP expression vector driven by the combined gene CXCL12-KDEL was constructed successfully, resulting in a good foundation block for the CXCL12-CXCR4 axis.