title: [Construction of recombinant replication competent plasmids containing HBV genome with mutations of precore and basic core promoter].
abstract: OBJECTIVE
  To construct a series of recombinant replication competent plasmids of hepatitis B virus (HBV) full-length genome with varying mutations of precore and basic core promoter.
  
  
  METHODS
  The plasmid containing the entire HBV (1.2 copies) genome was used to construct objective plasmids. The objective competent vectors were constructed by molecular cloning and PCR-based site-directed mutagenesis in vitro and confirmed with sequence analysis. After introducing the plasmids into hepatocellular carcinoma cell line (Huh7) by calcium phosphate transfection, the culture supernatant was collected to detect HBsAg and HBV DNA to analyze viral replication and expression.
  
  
  RESULTS
  Ten competent vectors with varying precore and basic core promoter mutations were constructed. After transfecting into Huh7, the vectors replicated and secreted viral particles.
  
  
  CONCLUSIONS
  Ten full-length competent HBV recombinants were obtained, which were harbouring varying mutations within precore and basic core promoter.
authors.affiliation: Institute of Hepatology, Peoples Hospital, Peking University, Beijing 100044, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.forename: Dong
authors.initials: D
authors.lastname: Jiang
authors.forename: Lai
authors.initials: L
authors.lastname: Wei
authors.forename: Hong-song
authors.initials: HS
authors.lastname: Chen
authors.forename: Xu
authors.initials: X
authors.lastname: Cong
authors.forename: Ran
authors.initials: R
authors.lastname: Fei
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D005822:Genetic Vectors; D016679:Genome, Viral; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014758:Viral Core Proteins / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
chemical_list: D006512:Hepatitis B Core Antigens; D011994:Recombinant Proteins; D014758:Viral Core Proteins