title: [Construction and expression of non-structural protein gene 3-4b of HCV 1b based on the adenoassociated virus vector].
abstract: OBJECTIVE
  To clone 1b type of HCV NS3-4b Gene and express in HEK 293 cells, lay the foundation for further study of the HCV NS3-4b recombinant adeno-associated virus vaccine and its dendritic cell vaccine.
  
  
  METHODS
  HCV 1b patients' serum was collected, and full length NS3-4b segment was amplified by RT-PCR and cloned into adeno-associated virus' expression vector pAAV. CMV. EGFP in order to express in HEK 293 cells. At last, it was validated whether express or not by Western Blot.
  
  
  RESULTS
  The 1b type gene NS3-4b were amplified and consistent to the expected size (2838 bp), the recombinant plasmid has been confirmed its successful restructured by double enzyme and sequencing, at last, Western Blot map can see objective protein expression after it transfect HEK 293 cells.
  
  
  CONCLUSIONS
  The adeno-associsted virus recombination HCV NS3-4b plasmid have successfully constructed and it can express in eukaryotic cells.
authors.affiliation: Department of Infection Disease, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China. lg3195@gmail.com
authors.forename: Tian
authors.initials: T
authors.lastname: Chen
authors.forename: Hai-Xia
authors.initials: HX
authors.lastname: Sun
authors.forename: Wei-Bin
authors.initials: WB
authors.lastname: Qin
authors.forename: Feng-Qin
authors.initials: FQ
authors.lastname: Zhu
authors.forename: Xue-Ling
authors.initials: XL
authors.lastname: Li
authors.forename: Hong
authors.initials: H
authors.lastname: Cao
authors.forename: Qi-Huan
authors.initials: QH
authors.lastname: Xu
authors.forename: Gang
authors.initials: G
authors.lastname: Li
mesh_terms: D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors; D057809:HEK293 Cells; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D010957:Plasmids; D014614:Vaccines, Synthetic / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000276:immunology; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D014614:Vaccines, Synthetic; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C080107:NS4B protein, flavivirus