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Synthesis of hepadnavirus particles that contain replication-defective duck hepatitis B virus genomes in cultured HuH7 cells.

Department of Human Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.

ABSTRACT

To evaluate the possibility of producing transducible replication-defective hepadnaviruses, cloned mutant duck hepatitis B virus genomes were tested both for virus antigen production and viral DNA synthesis following transfection into the human hepatoma cell line HuH7. Deletion of a cis-acting 12-nucleotide sequence implicated in viral DNA synthesis, direct repeat 1 (DR1), resulted in the loss of ability to synthesize both mature viral DNA and infectious virus. The delta DR1 mutant, however, produced envelope and core antigens and was shown to provide trans-acting functions required for the assembly of infection-competent particles. Thus, mutants with mutations in viral genes could be rescued as DNA-containing viral particles after cotransfection with delta DR1. The efficiency of rescue was influenced by the site of mutation. A mutant DNA encoding truncated core and envelope proteins not only was poorly rescued but also was able to suppress the production from a wild-type DNA of infectious virus.

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