title: [Construction and expression of dual promoter lentiviral vector containing TFR and VEGF genes].
abstract: OBJECTIVE
  To construct a dual promoter lentivira vector containing TFR and VEGF gene, and detect the expression of TFR and VEGF genes in MSCs of Chinese mini-swine.
  
  
  METHODS
  The TFR and VEGF gene were amplified by polymerase chain reaction and cloned into pLenti-GFP-Neo after the CMV and SV40 promoter respectively, to generate the lentivira vector pLenti-TG-VEGF. The four-plasmids lentiviral vector system(pRsv-REV, pMDlg-pRRE, pMD2G and pLenti-TG-VEGF)were cotransfected into human embryonic kidney 293 T cells with Lipofectin 2000 reagent. The packaged virus was harvested 72 h later. MSCs were infected by lentivirus carrying TFR and VEGF genes. Western blotting was used to detect the expression of TFR and VEGF genes in the viral infected MSCs.
  
  
  RESULTS
  Restriction endonuclease digestion analysis and DNA sequencing demonstrated that the dual promoter lentivirus vector containing TFR and VEGF genes were constructed successfully. MSCs could be successfully infected by the recombinant lentivirus. Expression of TFR and VEGF protein could be detected in the infected MSCs by Western blotting.
  
  
  CONCLUSIONS
  The dual promoter lentiviral vector containing TFR and VEGF genes are constructed successfully which provides basis for future research on Cardiac molecular imaging of cell transplantation.
authors.affiliation: Department of Radiology of Xijing Hospital, the Fourth Military Medical University, Xi'an 710032, China. weimengqi2008@163.com
authors.forename: Meng-qi
authors.initials: MQ
authors.lastname: Wei
authors.forename: Qi
authors.initials: Q
authors.lastname: Shen
authors.forename: Ying
authors.initials: Y
authors.lastname: Luo
authors.forename: Yi
authors.initials: Y
authors.lastname: Huan
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.forename: Yong
authors.initials: Y
authors.lastname: Yang
authors.forename: Jin-song
authors.initials: JS
authors.lastname: Zhang
authors.forename: Fan
authors.initials: F
authors.lastname: Lu
authors.forename: Min-wen
authors.initials: MW
authors.lastname: Zheng
mesh_terms: D000818:Animals; D015316:Genetic Therapy; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics*; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011990:Receptors, Transferrin / Q000235:genetics*; D013552:Swine; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
chemical_list: D011990:Receptors, Transferrin; D042461:Vascular Endothelial Growth Factor A