Aim: To create a model for studying ionic channels by means of the expressing human HCN2 and G418-resistant HEK293 cell lines established.
Methods: pcDNA3-hHCN2 was transfected with Lipofectin2000 into HEK293 cell line. The transfected cells would be survived in the further culture medium containing G418 antibiotic as the hHCN2 gene could express a G418 resistant products. Whole-cell patch clamp investigated that hHCN2 gene was transfected into HEK293 cells.
Results: The G418 resistant (600 ug/ml) HEK293 cell line was established successfully and whole-cell patch clamp recorded ionic currents of transfected hHCN2.
Conclusion: The G418 resistant HEK293 cell line was successfully established with transfection of plasmid pcDNA3-hHCN2 by Lipofectin, which might be useful for studying the relationship between the structure and function of cloned ionic channels.