Establishment of a HEK293 cell line stably expressing human HCN2 gene.
- Author:
Xin ZHAO
1
;
Xiang-Jun YANG
;
Xia BAI
;
Hong-Xia LI
;
Xu-Jie CHENG
;
Wen-Ping JIANG
Author Information
- Publication Type:Journal Article
- MeSH: Gene Expression; Genetic Vectors; HEK293 Cells; Humans; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; Ion Channels; genetics; Patch-Clamp Techniques; Plasmids; Potassium Channels; Transfection
- From: Chinese Journal of Applied Physiology 2006;22(2):254-256
- CountryChina
- Language:Chinese
-
Abstract:
AIMTo create a model for studying ionic channels by means of the expressing human HCN2 and G418-resistant HEK293 cell lines established.
METHODSpcDNA3-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.
RESULTSThe G418 resistant (600 ug/ml) HEK293 cell line was established successfully and whole-cell patch clamp recorded ionic currents of transfected hHCN2.
CONCLUSIONThe 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.
