Construction and expression of ATP50 fluorescent protein in TM3 mouse Leydig cells.
- Author:
Liang CHEN
1
;
Zhong-cheng XIN
;
Xue-jun JIANG
;
Long TIAN
;
Yi-ming YUAN
;
Gang LIU
;
Wei-dong SONG
;
Ying-lu GUO
Author Information
- Publication Type:Journal Article
- MeSH: Adenosine Triphosphatases; biosynthesis; genetics; Animals; Carrier Proteins; biosynthesis; genetics; Cells, Cultured; Cloning, Molecular; Genetic Vectors; Green Fluorescent Proteins; biosynthesis; genetics; Leydig Cells; metabolism; Male; Membrane Proteins; biosynthesis; genetics; Mice; Mitochondria; metabolism; Recombinant Fusion Proteins; biosynthesis; Transfection
- From: National Journal of Andrology 2006;12(11):992-996
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the expression and localization of ATP50 by construction of ATP50-pEYFP-N1 in primary cultured mouse Leydig cells.
METHODSPrimary cultured mouse Leydig cells were confirmed by 3B-HSD staining. ATP50 was cloned into pEYFP-N1 between Bam HI and Eco RI sites. Cell-transfection and living-cell fluorescence imaging microscopy were employed to investigate the sub-cellular localization of YFP-ATP50 in TM3 mouse Leydig cells.
RESULTSATP50 green fluorescent protein was well co-localized with red fluorescence mitochondrion marker-Mitotracker in TM3 mouse Leydig cells.
CONCLUSIONATP50 was expressed in primary cultured mouse Leydig cells. The fluorescent expression vector of ATP50 was constructed successfully and YFP-ATP50 was located in mitochondria in TM3 mouse Leydig cells, which provided a useful clue for further research on the steroidogenesis dysfunction in aging males.
