title: [Construction and expression of the efficient recombinant plasmid pEE14.1-IFN-α expressing human IFN-α].
abstract: OBJECTIVE
  To construct the eukaryotic expression plasmid pEE14.1-IFN-α expressing human IFN-α gene,  and to detect the expression of the plasmid in eukaryotic cells.
  
  
  METHODS
  The human IFN-α gene amplified by  PCR  and   was linked into pCI-GPI,   then inserted into the eukaryotic expression vector pEE14.1. The recombinant plasmid was transfected into the 293T cells,    the  IFN-α   expression was detected by ELISA and Western blotting.
  
  
  RESULTS
  Enzyme digestion and sequence analysis showed that the bicistronic eukaryotic expression vector pEE14.1-IFN-α was constructed successfully. The expression of plasmid was detected by ELISA,   and the production of IFN-α in supernatant of transfected cells was about 3.15 ng/mL. Also,    Western blotting could reveal the characteristic band of IFN-α gene.
  
  
  CONCLUSIONS
  The vector is constructed successfully,   which provide a new selection for HBV immunotherapy.
authors.affiliation: College of Life Science and Technology,    Qufu normal University,    Qufu  273165,    China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Gao
authors.forename: Xi
authors.initials: X
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Yuan-Ji
authors.initials: YJ
authors.lastname: Xu
authors.forename: Ji-Yun
authors.initials: JY
authors.lastname: Yu
authors.forename: Bing-Ran
authors.initials: BR
authors.lastname: Guo
authors.forename: Jin-Qi
authors.initials: JQ
authors.lastname: Yan
mesh_terms: D015153:Blotting, Western; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D016898:Interferon-alpha / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins; D004247:DNA