Abstract:GIP receptor agonists which can stimulate insulin secretion could be potential drugs in the treatment of type II diabetes. Therefore, it is important of the screening model for GIP receptor agonists. In this study, GIPR gene was amplified by PCR, digested by HindIII/XhoI and then connected to the expression vector pCMV6ACGFP to construct pCMV6ACGIPRGFP recombinant plasmid. The recombinant plasmid was transformed into Escherichia coli DH5α, subsequently confirmed by colony PCR, restriction endonuclease digestion and sequencing. The recombinant plasmid was transfected into RINm5F cells by lipofectamine 2000. After screening with G418, the monoclonal RINm5F/GIPRGFP cell line was selected. The results showed that the GIPR gene with a length of
1 319 bp was amplified and cloned into the eukaryotic expression vector of pCMV6ACGFP. After analysis of colony PCR, enzyme digestion and sequence, the construction of plasmid pCMV6ACGIPRGFP was proved to be successful. Lots of positive puncta were observed in the treatment of GIPR agonist of (DAla2) GIP .The results reveal that GIPRGFP cell line with stable expression was successfully constructed. This cell line can be used to screen agonists of GIPR for exploring potential new medicines for diabetes.