title: [Construction and expression of eukaryotic expression plasmid pIRES-neo-HBAg carrying HBV fusion antigen gene].
abstract: OBJECTIVE
  To construct a eukaryotic expression plasmid harboring HBV fusion antigen gene, and to express it in 293T cells.
  
  
  METHODS
  The HBV fusion gene fragment was amplified by PCR from the plasmid pVAX1-HBV containing HBV fusion gene. After purified, the product was cloned into pMD18-T vector. The recombinant plasmid was confirmed by endonuclease digestion and sequencing analysis, and then subcloned into eukaryotic expression vector pIRES-neo. Then the recombinant expression plasmid pIRES-neo-HBAg was transfered into 293T cells by Lipofectamine(TM); 2000. The expression of HBV fusion antigen was identified by Western blotting, flow cytometry and immunofluorescence cytochemistry.
  
  
  RESULTS
  The eukaryotic expression vector pIRES-neo-HBAg was constructed successfully. The expression of fusion antigen could be detected in the pIRES-neo-HBAg transfected 293T cells by Western blotting, immunofluorescence cytochemistry and flow cytometry.
  
  
  CONCLUSIONS
  The eukaryotic expression plasmid pIRES-neo-HBAg is successfully constructed and the fusion antigen is expressed in 293T cells.
authors: Cell Engineering Research Center, Fourth Military Medical University, Xi'an 710032, China. drlinwang@163.com
authors: Lin
authors: L
authors: Wang
authors: Yu
authors: Y
authors: Wang
authors: Hao
authors: H
authors: Wu
authors: Xiaoming
authors: X
authors: Zhu
authors: Jiyun
authors: J
authors: Yu
authors: Jianli
authors: J
authors: Jiang
authors: Jinqi
authors: J
authors: Yan
mesh_terms: D002460:Cell Line; D003001:Cloning, Molecular; D005434:Flow Cytometry; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006511:Hepatitis B Antigens / Q000235:genetics* / Q000276:immunology; D006801:Humans; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D014162:Transfection
chemical_list: D006511:Hepatitis B Antigens; D011993:Recombinant Fusion Proteins