Adeno-associated virus DNA replication in vitro: activation by a maltose binding protein/Rep 68 fusion protein.

摘要: The adeno-associated virus (AAV) nonstructural protein Rep 68 is required for viral DNA replication. An in vitro assay has been developed which addition of to an extract from uninfected HeLa cells supports AAV In this paper, we report characterization the replication process when a fusion maltose binding and 68, expressed Escherichia coli, was used assay. Replication observed template either linear double-stranded or plasmid construct containing intact DNA. When recombinant as template, there pBR322 well DNA; however, not replicated. appeared initiate on led separation sequences those vector, termed rescue. There no evidence that could products can make site-specific nick 124 nucleotides 3' end site called terminal resolution site. Our data are most consistent with initiation occurring at proceeding toward terminus. construct, elongation continued past junction into newly synthesized sequence hairpinned, switched strands, replicated 4 h, during time 70% maximal synthesis took place. additional finding resolve dimer length duplex intermediates monomer duplexes without synthesis.