CONSTRUCTION OF RECOMBINANT hNaDC3/GFP AND ITS LOCALIZATION IN RENAL PROXIMAL TUBULE EPITHELIA

To construct the fusion gene of hNaDC3 with green fluorescent protein (GFP), and study the localization of the fusion protein in renal proximal tuble epithelia, the full length cDNA of hNaDC3 was amplified by RT-PCR from normal human renal tissues, and then cloned into pGEM-T vector. After DNA sequencing, the hNaDC3 was inserted into mammalian expression plasmid pEGFP-C3 successfully. The fused gene was then transfected into LLC-PK1 cell with lipofectamine. Confocal fluorescence microscopy showed that green fluorescence spread over the whole cell transfected with pEGFP-C3. After transfection with pEGFP-C3-hNaDC3, fluorescence distributed in both the cytoplasm and on the plasma membrane first day, located mainly on the plasma membrane fifth day. It is suggested that the hNaDC3 protein localizes on the plasma membrane.