title: [Construction and identification of luciferase and EGFP expression vectors containing human IFN-β promoter].
abstract: OBJECTIVE
  To construct a luciferase expression vector (pGL3-IFNB1) and an enhanced green fluorescent protein expression vector (pGE3-IFNB1) containing human IFN-β promoter, and verify the promoter activity of IFN-β in A549 cells.
  
  
  METHODS
  IFN-β promoter was amplified with the human genome DNA by PCR, then the segment was cloned into the eukaryotic expression vectors pGL3-basic and pGE3-basic by PCR. The eukaryotic expression vectors were named pGL3-IFNB1 and pGE3-IFNB1, respectively. The recombinant vectors were then transiently transfected into A549 cells respectively, and 6 hours later, the transfected cells were infected with Hantaan virus (HTNV). After 24 hours, the expressions of pGL3-IFNB1 and pGE3-IFNB1 were tested in A549 cells.
  
  
  RESULTS
  Double restriction enzyme digestion and sequence analysis showed that the recombinant vectors were successfully constructed and expressed in A549 cells correctly. After HTNV infection, stronger expression was observed in these cells.
  
  
  CONCLUSIONS
  The recombinant vectors pGL3-IFNB1 and pGE3-IFNB1 containing IFN-β promoter have been successfully constructed. It provides useful tools for further study on the mechanism of IFN-β production induced by virus.
authors.affiliation: Department of Microbiology, Fourth Military Medical University, Xi'an 710032; Laboratory, No. 323 Hospital of PLA, Xi'an 710054, China.
authors.forename: Yongni
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Mengyuan
authors.initials: M
authors.lastname: Cao
authors.affiliation: Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Qianqian
authors.initials: Q
authors.lastname: Hui
authors.affiliation: Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Ye
authors.affiliation: Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Yu
authors.affiliation: Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Fanglin
authors.initials: F
authors.lastname: Zhang
mesh_terms: D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D018063:Hantaan virus / Q000502:physiology; D054884:Host-Pathogen Interactions; D006801:Humans; D016899:Interferon-beta / Q000235:genetics*; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009369:Neoplasms / Q000235:genetics / Q000473:pathology / Q000821:virology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins; D016899:Interferon-beta; D008156:Luciferases