title: [Construction and expression of ATP50 fluorescent protein in TM3 mouse Leydig cells].
abstract: 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.
authors.affiliation: Andrology Center, First Hospital, Peking University, Beijing 100009, China. bdyychenliang@163.com
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
authors.forename: Zhong-cheng
authors.initials: ZC
authors.lastname: Xin
authors.forename: Xue-jun
authors.initials: XJ
authors.lastname: Jiang
authors.forename: Long
authors.initials: L
authors.lastname: Tian
authors.forename: Yi-ming
authors.initials: YM
authors.lastname: Yuan
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.forename: Wei-dong
authors.initials: WD
authors.lastname: Song
authors.forename: Ying-lu
authors.initials: YL
authors.lastname: Guo
mesh_terms: 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
chemical_list: 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