title: [Effects of nuclear factor of activated T cells  on the promoter activity of constitutively activated SV40].
abstract: OBJECTIVE
  To check   if  the common used constitutively promoters,  such as CMV,  TK and SV40 could  be  responded  to the nuclear factor of activated T cell (NFAT),  and to explore the strategies to choose rational internal control in the dual luciferase reporter assay.
  
  
  METHODS
  pCMV-luc vector,  in which luciferase activity is driven by CMV promoter,  was cloned by amplifying the CMV promoter fragment from the pCDNA3.1 vector and then inserting the CMV promoter region into the pGL3-basic vector using the standard protocol. pTK-Luc reporter was similarly constructed,  with the TK promoter from the pRL-TK vector. The constructed pCMV-Luc or pTK-Luc was co-transfected with pBIND or pRL-TK respectively,  together with NFAT or constitutively active form named NFATCA. Relative luciferase activity was calculated as instructed by the manual instruction.
  
  
  RESULTS
  Both pCMV-Luc and pTK-Luc vectors were successfully constructed. Luciferase activity assay revealed that SV40 promoter responded to active NFAT.
  
  
  CONCLUSIONS
  The common used internal control promoter SV40 could respond to active NFAT,  which should be kept in mind for selection of the rational internal control vector in the dual luciferase reporter assay. In addition,  our study here also provides a practical strategy for rational selection of the internal control.
authors.affiliation: Department of General Surgery,  Tangdu Hospital of the Fourth Military Medical University,  Xi'an   710038,  China.
authors.forename: Jun
authors.initials: J
authors.lastname: Pu
authors.forename: Guo-dong
authors.initials: GD
authors.lastname: Yang
authors.forename: Xiao-zhao
authors.initials: XZ
authors.lastname: Lu
authors.forename: Meng-ying
authors.initials: MY
authors.lastname: Wei
authors.forename: Zi-fan
authors.initials: ZF
authors.lastname: Lu
authors.forename: Jian-guo
authors.initials: JG
authors.lastname: Lu
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D050778:NFATC Transcription Factors / Q000502:physiology*; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics*
chemical_list: D050778:NFATC Transcription Factors; D008156:Luciferases