Construction of eukaryotic expression vectors for rat cardiac kir2.2, kir2.3 channels and their expressions in HEK293 cells

Objective To construct eukaryotic expression vectors for rat cardiac kir2.2,kir2.3 channels and to determine their expressions in HEK293 cells.Methods Rat cardiac orthologs of Kir2.2 and Kir2.3 were cloned by reverse transcriptase-PCR and were subcloned into the eukaryotic expression vector pEGFP-N1.The constructed recombinant plasmids pEGFP-N1-kir2.2 and pEGFP-N1-kir2.3 were transfected into human embryonic kidney-293(HEK293)cells.The currents of kir2.2 and kir2.3 channels were identified by whole-cell patch-clamp technique.Results Restriction analysis and sequencing proved that the recombinant plasmids pEGFP-N1-kir2.2 and pEGFP-N1-kir2.3 were constructed correctly. The Kir2.2 and Kir2.3 channels were successfully expressed in HEK293 cells and Kir2.2 current(I_Kir2.2)and Kir2.3 current(I_Kir2.3)were recorded.Conclusion The eukaryotic expression vectors for rat cardiac kir2.2 and kir2.3 channels were successfully constructed and expressed in HEK293 cells.