title: [Construction and expression of eukaryotic expression vector for pEGFP-GPC3].
abstract: OBJECTIVE
  To construct and expression of eukaryotic  expression vector of glypican-3 (GPC3) pEGFP-GPC3 in mouse dendritic cells (DCs).
  
  
  METHODS
  The target gene fragment of GPC3 and empty vector were obtained from plasmid pCMV-SPORT6-GPC3 and vector pEGFP-N1  digested by double enzymes. GPC3 and empty vector were linked by ligase T4,  and then transfected into E.coli DH5α. Identification with EcoRI,  BamHI digestion,  the recombinant eukaryotic  expression vector pEGFP-GPC3 was transfected into DCs by Lipofectamine(TM);2000. Fluorescent microscopy and Western blot were used to detected the expression of GPC3.
  
  
  RESULTS
  The expression of EGFP-GPC3 fusion protein  was detected in DCs,  and GPC3 fusion protein of 67   kDa  was detected by Western blotting.
  
  
  CONCLUSIONS
  The eukaryotic expression vector pEGFP-of GPC3 was constructed succesfully and transfected  effectively into DCs.
authors.affiliation: Department of Hepatobiliary Surgery, Guangxi Medical University, Nanning, China.
authors.forename: Chun-hui
authors.initials: CH
authors.lastname: Liu
authors.forename: Zhi-ming
authors.initials: ZM
authors.lastname: Zhang
authors.forename: Jian-yong
authors.initials: JY
authors.lastname: Liu
mesh_terms: D000818:Animals; D003713:Dendritic Cells / Q000378:metabolism; D005822:Genetic Vectors; D053673:Glypicans / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D014162:Transfection
chemical_list: D053673:Glypicans; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins