title: Development of a Hepatitis B Virus Reporter System to Monitor the Early Stages of the Replication Cycle.
abstract: Currently, it is possible to construct recombinant forms of various viruses, such as human immunodeficiency virus 1 (HIV-1) and hepatitis C virus (HCV), that carry foreign genes such as a reporter or marker protein in their genomes. These recombinant viruses usually faithfully mimic the life cycle of the original virus in infected cells and exhibit the same host range dependence. The development of a recombinant virus enables the efficient screening of inhibitors and the identification of specific host factors. However, to date the construction of recombinant hepatitis B virus (HBV) has been difficult because of various experimental limitations. The main limitation is the compact genome size of HBV, and a fairly strict genome size that does not exceed 1.3 genome sizes, that must be packaged into virions. Thus, the size of a foreign gene to be inserted should be smaller than 0.4 kb if no deletion of the genome DNA is to be performed. Therefore, to overcome this size limitation, the deletion of some HBV DNA is required. Here, we report the construction of recombinant HBV encoding a reporter gene to monitor the early stage of the HBV replication cycle by replacing part of the HBV core-coding region with the reporter gene by deleting part of the HBV pol coding region. Detection of recombinant HBV infection, monitored by the reporter activity, was highly sensitive and less expensive than detection using the currently available conventional methods to evaluate HBV infection. This system will be useful for a number of applications including high-throughput screening for the identification of anti-HBV inhibitors, host factors and virus-susceptible cells.
authors.affiliation: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine; lbnishitsuji@hospk.ncgm.go.jp.
authors.forename: Hironori
authors.initials: H
authors.lastname: Nishitsuji
authors.affiliation: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Yamamoto
authors.affiliation: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine.
authors.forename: Ritsuko
authors.initials: R
authors.lastname: Shiina
authors.affiliation: Central Pharmaceutical Research Institute, Japan Tobacco Inc.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Harada
authors.affiliation: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine.
authors.forename: Saneyuki
authors.initials: S
authors.lastname: Ujino
authors.affiliation: Research Center for Hepatitis and Immunology, National Center for Global Health and Medicine.
authors.forename: Kunitada
authors.initials: K
authors.lastname: Shimotohno
mesh_terms: D004279:DNA, Viral; D017930:Genes, Reporter / Q000502:physiology*; D016679:Genome, Viral / Q000502:physiology; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000254:growth & development / Q000502:physiology*; D006801:Humans; D014162:Transfection; D014771:Virion / Q000502:physiology; D014779:Virus Replication / Q000502:physiology*
chemical_list: D004279:DNA, Viral