title: [Preparation of green fluorescent protein retrovirus and its application in mediating gene transfer into retinal pigment epithelial cells].
abstract: OBJECTIVE
  To determine whether human retinal pigment epithelial (RPE) cells can be infected by retrovirus and modified by retrovirus-mediated gene transfer.
  
  
  METHODS
  Recombination retroviral vector pLNCX-GFP (green fluorescent protein, GFP) was generated by inserting 780 bp GFP cDNA fragment into the MCS site of pLNCX. pLNCX-GFP was transfected into ecotropic packaging cell line PhiX-Eco and amphotropic packaging cell line PhiX-Ampho and PA317. Retroviral titer was tested by counting GFP expression of NIH3T3 cells. Then RPE cells were infected by using GFP retrovirus-containing supernatant.
  
  
  RESULTS
  GFP was expressed and retrovirus was produced upon pLNCX-GFP being transfected into packaging cell line. The GFP retrovirus was able to infect primary cultured human RPE cells and immortalized RPE cell line.
  
  
  CONCLUSIONS
  The retrovirus can introduce a foreign gene into RPE cells efficiently, thereby it can be used as an important tool to deliver gene into RPE for therapy of fundus diseases.
authors.affiliation: Central Laboratory, The First People's Hospital of Shanghai, Shanghai 200080, China.
authors.forename: Q
authors.initials: Q
authors.lastname: Huang
authors.forename: P
authors.initials: P
authors.lastname: Xu
authors.forename: W
authors.initials: W
authors.lastname: Liu
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.forename: Q
authors.initials: Q
authors.lastname: Gu
authors.forename: S
authors.initials: S
authors.lastname: Tian
authors.forename: Y
authors.initials: Y
authors.lastname: Fan
authors.forename: K
authors.initials: K
authors.lastname: Xig
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.forename: C
authors.initials: C
authors.lastname: Li
mesh_terms: D002478:Cells, Cultured; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D010857:Pigment Epithelium of Eye / Q000166:cytology*; D012190:Retroviridae / Q000235:genetics*
chemical_list: D049452:Green Fluorescent Proteins