In vitro and in vivo replication of a chemically synthesized consensus genome of hepatitis B virus genotype B

J Virol Methods. 2015 Mar:213:57-64. doi: 10.1016/j.jviromet.2014.11.007. Epub 2014 Nov 27.

Abstract

Hepatitis B virus (HBV) genotypes vary in their geographical distribution and virological features. Previous investigations have shown that HBV genotype B is a predominant HBV genotype in China. Studies on HBV concerning different isolates frequently meet the question about the HBV reference strain and its representativeness. Although HBV consensus sequences can be generated easily by sequence alignment, they may not exist in nature or could not usually be isolated from patient samples. Thus, the construction of a consensus HBV genome has been proposed. In this study, an HBV genotype B consensus sequence was established by comparing 42 full-length HBV genotype B sequences and the genome was generated by chemical synthesis. This consensus genome was fully replication competent by in vitro transfection into hepatoma cells. The plasmid pHBV1.3B carrying a 1.3× full-length HBV consensus genome was hydrodynamically injected into Balb/c mice. HBsAg, anti-HBs, HBeAg, anti-HBe, and anti-HBc detection indicated expression and replication of this HBV genome in mice, similar to other HBV isolates. This approach represents a strategy to design and create consensus HBV genomes for future studies.

Keywords: Consensus genome; Genotype; Hepatitis B virus; Replication.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Consensus Sequence
  • DNA, Viral / chemical synthesis*
  • DNA, Viral / genetics
  • Genome, Viral*
  • Genotype
  • Hepatitis B Antibodies / blood
  • Hepatitis B Surface Antigens / blood
  • Hepatitis B virus / genetics
  • Hepatitis B virus / physiology*
  • Hepatocytes / virology
  • Humans
  • Male
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Sequence Analysis, DNA
  • Virus Replication*

Substances

  • DNA, Viral
  • Hepatitis B Antibodies
  • Hepatitis B Surface Antigens

Associated data

  • GENBANK/GQ205440