Abstract:
To explore the function of TRPM7 (transient receptor potential melastatin 7) channel,the overexpression system of TRPM7 channel was constructed by transiently transfection of HEK293T (human embryonic kidney 293T) cell with TRPM7-eGFP plasmid.Through the fluorescence microscope analysis,the transfection efficiency was about 30%-40%.The corresponding expression of the gene and protein was observed by RT-PCR and western blotting. And the result of patch clamp showed that the whole cell current was about 2800pA.Acidic extracellular solution increased the TRPM7 inward currents and 200μmol/L 2-APB(2-aminoethyl diphenyl borate) inhibited the TRPM7 outward currents.These results demonstrated that the transiently transfected TRPM7-HEK293T cell line was successfully constructed.The acidic pH and the non-specific inhibitor of TRPM7,2-APB made different changes on TRPM7,which indicates that the function of the channel can be affected by multiple factors.