title: [Stable expression of human thyrotropin receptor in HEK 293T cells].
abstract: OBJECTIVE
  To establish a strain of eukaryotic cells with overexpressed human thyrotropin receptor (hTSHR) for the development of anti-thyroid drugs.
  
  
  METHODS
  GV266 vector and hTSHR gene were digested by Age I/Nhe I. Plasmids expressing GV266-hTSHR were constructed using T4 ligase and then transfected into 293T cells. The expression of hTSHR was determined by Western blot. Packaging plasmids were built in the 293T cells with Opti-MEM, Lipofectamine 2000 and helper plasmid. The titer of the packaging plasmids was determined with qPCR. The packaging plasmids and the plasmids expressing GV266-hTSHR were co-transfected into 293T cells to obtain a strain of cells (GV266-hTSHR-293T) with stable expression of hTSHR. The GV266-hTSHR293T stain was detected by green fluorescent protein (GFP) fluorescence.
  
  
  RESULTS
  The DNA sequence of GV266-hTSHR matched that of hTSHR. The Western blot showed a 62 x 10(3) target band. The titer of packaging plasmids reached 2 x10(8) TU/mL. The GV266-hTSHR-293T cells were visible under GFP fluorescence.
  
  
  CONCLUSIONS
  HEK 293T cells with stable expression of hTSHR was established.
authors: Department of Nuclear Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
authors: Gong-Shun
authors: GS
authors: Tang
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D057809:HEK293 Cells; D006801:Humans; D010957:Plasmids; D011989:Receptors, Thyrotropin / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D011989:Receptors, Thyrotropin; D049452:Green Fluorescent Proteins