title: [Construction and expression of the recombinant plasmid containing BddhFVIII in HepG2 cells].
abstract: OBJECTIVE
  To get stable cell line expressing B domain-deleted human FVIII (BDDhFVIII) by constructing the eukaryotic expression plasmid.
  
  
  METHODS
  Eukaryotic expression plasmid containing BDDhFVIII was constructed and transfected into HepG2 cells via electroporation. The expression and purification of the target protein was detected by Western blot.
  
  
  RESULTS
  Results of enzyme digestion and sequence analysis demonstrated that the gene of BDDhFVIII was correctly inserted into the eukaryotic expression vector pcDNA4/v5-his. Western blot confirmed the successful expression of BDDhFVIII at the protein levels in HepG2 cells.
  
  
  CONCLUSIONS
  The constructed eukaryotic expression vector was able to generate high level expression of human FVIII in HepG2 cells, thus could construct human blood coagulation FVIII stable cell line successfully.
authors.affiliation: Department of Clinical Laboratory Science, Tianjin Medical University, Tianjin, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhao
authors.forename: Jin-hui
authors.initials: JH
authors.lastname: Xie
authors.forename: Shuang-yu
authors.initials: SY
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Dong
authors.forename: Jing-hui
authors.initials: JH
authors.lastname: Chong
authors.forename: Li-na
authors.initials: LN
authors.lastname: Yan
authors.forename: Yun-de
authors.initials: YD
authors.lastname: Liu
authors.forename: Yu-hua
authors.initials: YH
authors.lastname: Yuan
mesh_terms: D018274:Electroporation; D005169:Factor VIII / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000096:biosynthesis*; D006467:Hemophilia A / Q000235:genetics; D056945:Hep G2 Cells; D006801:Humans; D010957:Plasmids / Q000096:biosynthesis*; D011995:Recombination, Genetic
chemical_list: D005169:Factor VIII