[Construction and expression of ATP50 fluorescent protein in TM3 mouse Leydig cells].
OBJECTIVE
To study the expression and localization of ATP50 by construction of ATP50-pEYFP-N1 in primary cultured mouse Leydig cells.


METHODS
Primary 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.


RESULTS
ATP50 green fluorescent protein was well co-localized with red fluorescence mitochondrion marker-Mitotracker in TM3 mouse Leydig cells.


CONCLUSIONS
ATP50 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.
Chen
Liang
L
Andrology Center, First Hospital, Peking University, Beijing 100009, China. bdyychenliang@163.com
Xin
Zhong-cheng
ZC
Jiang
Xue-jun
XJ
Tian
Long
L
Yuan
Yi-ming
YM
Liu
Gang
G
Song
Wei-dong
WD
Guo
Ying-lu
YL
D000251:Adenosine Triphosphatases / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D007985:Leydig Cells / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D025261:Mitochondrial Proton-Translocating ATPases; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection
D002352:Carrier Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D000251:Adenosine Triphosphatases; D025261:Mitochondrial Proton-Translocating ATPases; C020439:oligomycin sensitivity-conferring protein