title: Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA.
abstract: OBJECTIVE
  To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs).
  
  
  METHODS
  Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19 x 10(12) copies of linear HBV DNA/1 x 10(7) PDHs). After 1-5 d of transfection, HBsAg and HBeAg in the supernatant and lysate of PDHs were measured with the IMX System. Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PDHs electroporated were used as control group.
  
  
  RESULTS
  HBsAg in the hepatocyte lysates of transfected group was 15.24 (1 d), 14.55 (3 d) and 5.13 (5 d; P/N values, positive > or =2.1) respectively. HBeAg was negative (<2.1). Both HBsAg and HBeAg were negative in the supernatant of transfected group. Dot blotting revealed that HBV DNA was strongly positive in the transfected group and negative in the control group. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (rc DNA), covalently closed circular (ccc DNA), and single-stranded (ss DNA) HBV DNA replicative intermediates in the transfected group, there was no integrated HBV DNA in the cellular genome. These parameters were negative in control group.
  
  
  CONCLUSIONS
  Expression and replication of HBV genes can occur in hepatocytes from non-mammalian species. HBV replication has no critical species-specificity, and yet hepatic-specific regulating factors in hepatocytes may be essential for viral replication.
authors.affiliation: Department of Infectious Diseases of the First Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400016, China. cqyaoyunqing@126.com
authors.forename: Yun-Qing
authors.initials: YQ
authors.lastname: Yao
authors.forename: Ding-Feng
authors.initials: DF
authors.lastname: Zhang
authors.forename: Ni
authors.initials: N
authors.lastname: Tang
authors.forename: Ai-Long
authors.initials: AL
authors.lastname: Huang
authors.forename: Xiao-Yi
authors.initials: XY
authors.lastname: Zou
authors.forename: Jiang-Feng
authors.initials: JF
authors.lastname: Xiao
authors.forename: Yun
authors.initials: Y
authors.lastname: Luo
authors.forename: Da-Zhi
authors.initials: DZ
authors.lastname: Zhang
authors.forename: Bo
authors.initials: B
authors.lastname: Wang
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Zhou
authors.forename: Hong
authors.initials: H
authors.lastname: Ren
authors.forename: Qi
authors.initials: Q
authors.lastname: Liu
authors.forename: Shu-Hua
authors.initials: SH
authors.lastname: Guo
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics; D004372:Ducks; D018347:Hepadnaviridae Infections / Q000821:virology*; D015611:Hepatitis B Virus, Duck / Q000235:genetics* / Q000254:growth & development*; D006524:Hepatitis, Viral, Animal / Q000821:virology*; D022781:Hepatocytes / Q000166:cytology / Q000821:virology*; D014162:Transfection; D014779:Virus Replication
chemical_list: D004279:DNA, Viral