title: [Cloning of human Nanog gene and its expression in COS-7L cells].
abstract: OBJECTIVE
  To clone human Nanog gene and express it in mammalian cells.
  
  
  METHODS
  LA-PCR was used to amplify human Nanog gene from genomic DNA of HEK 293 cells. The Nanog gene was inserted into the Flag-tagged pCMV vector by gene recombination technique. After being confirmed by sequencing, the pFlag-Nanog was transfected into COS-7L cells. Western blot and indirect immunofluorescent staining against the Flag tag were used to detect the expression of Nanog protein.
  
  
  RESULTS
  The sequence of the cloned Nanog was confirmed to be correct by sequence analysis. The COS-7L cells transfected with pFlag-Nanog could efficiently express Nanog protein.
  
  
  CONCLUSIONS
  The cloning of human Nanog gene and the construction of its eukaryotic expression vector were successful. The study will lay the foundation for further research on the function of Nanog and its role in neural stem cells.
authors.affiliation: Institute of Neuroscience, Fourth Military Medical University, Xi'an 710032, China. Liu123xia5798@sohu.com
authors.forename: Xia
authors.initials: X
authors.lastname: Liu
authors.forename: Mei
authors.initials: M
authors.lastname: Long
authors.forename: Hui
authors.initials: H
authors.lastname: Jiang
authors.forename: Wei-lin
authors.initials: WL
authors.lastname: Jin
authors.forename: Xi-ying
authors.initials: XY
authors.lastname: Jiao
authors.forename: Gong
authors.initials: G
authors.lastname: Ju
mesh_terms: D000818:Animals; D019556:COS Cells / Q000378:metabolism; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics*; D018398:Homeodomain Proteins / Q000096:biosynthesis* / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000166:cytology; D000071317:Nanog Homeobox Protein; D017421:Sequence Analysis
chemical_list: D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D004247:DNA