Establishment of Na+-glucose cotransporter 2 heterologous expression system in HEK293 cells
10.3760/cma.j.issn.1001-7097.2011.08.012
- VernacularTitle:人胚肾细胞SGLT2基因异源表达体系的构建
- Author:
Lei YU
;
Xujie ZHOU
;
Jicheng LV
;
Jiaxiang DING
;
Li ZHU
;
Hong ZHANG
- Publication Type:Journal Article
- Keywords:
Sodium-glucose transporter 2;
Glycosuria,renal;
Transfection;
Familial renal glucosuria;
Heterologous expression system
- From:
Chinese Journal of Nephrology
2011;27(8):606-610
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish heterologous expression system of Na+-glucose cotransporter 2 (SGLT2) gene.Methods Human SGLT2 cDNA from normal kidney, generated by RT-PCR,was subclonedintoPEXL-GFP vector andtransfectedinto HEK293cells. After 24hours of incubation, the expression of SGLT2-GFP fusion protein was detected by Western blotting and laser confocal microscopy.Transport activity of SGLT2-GFP fusion proteins in cultured human HEK293 cells was evaluated with the uptake test of glucose analogue.ResultsSGLT2-GFP fusion protein was expressed in cultured human HEK293 cells.Furthermore, confocal microscopy using green fluorescent protein(GFP) revealed a punctate membrane pattern of SGLT2.Glucose analogue uptake increased in HEK293 cells transfected with SGLT2-GFP at least by 3.5 folds compared with HEK293 cells transfected with GFP vector only(P＜0.01).Conclusion Heterologous expression of SGLT2 gene in HEK293 cells is successfully established, which provides valuable approach for the functional and pathological study of SGLT2 gene.
