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Although HBV polymerase (HBVpol, pol) plays a pivotal role in HBV replication and must be a definite therapeutic target. The problems are that the detailed functions and intracellular dynamics of HBVpol remain unclear. Here, we constructed two kinds of tagged HBVpol, PA\u2010tagged and HiBiT\u2010tagged pol, and the HBV\u2010producing vectors. Each PA tag and HiBiT tag were inserted into N\u2010terminus of spacer region on HBVpol open reading frame. Transfection of the plasmids into HepG2 cells led to production of HBV. These tagged HBVpol were detectable in HBV replicating cells and pol\u2010HiBiT enabled quantitative analysis. Furthermore, these recombinant HBV were infectious to primary human hepatocytes. Thus, we successfully designed infectious and replication\u2010competent recombinant HBV harboring detectable tagged HBVpol. Such infectious recombinant HBV will provide a novel tool to study HBVpol dynamics and develop new therapeutics against HBV.<\/jats:p>","DOI":"10.1111\/1348-0421.13183","type":"journal-article","created":{"date-parts":[[2024,12,2]],"date-time":"2024-12-02T11:27:50Z","timestamp":1733138870000},"page":"43-58","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Generation of Replication\u2010Competent Hepatitis B Virus Harboring Tagged Polymerase for Visualization and Quantification of the Infection"],"prefix":"10.1111","volume":"69","author":[{"given":"Chiharu","family":"Morita","sequence":"first","affiliation":[{"name":"Division of Virology, Department of Microbiology and Immunology Osaka University Graduate School of Medicine Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masami","family":"Wada","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Microbiology and Immunology Osaka University Graduate School of Medicine Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1496-0964","authenticated-orcid":false,"given":"Eriko","family":"Ohsaki","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Microbiology and Immunology Osaka University Graduate School of Medicine Osaka Japan"},{"name":"Center for Infectious Disease Education and Research (CiDER) Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shihoko","family":"Kimura\u2010Ohba","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Microbiology and Immunology Osaka University Graduate School of Medicine Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5641-8105","authenticated-orcid":false,"given":"Keiji","family":"Ueda","sequence":"additional","affiliation":[{"name":"Division of Virology, Department of Microbiology and Immunology Osaka University Graduate School of Medicine Osaka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,12,2]]},"reference":[{"key":"e_1_2_11_2_1","unstructured":"WHO. 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