Systematic Excision of Vector Sequences from the BAC-Cloned Herpesvirus Genome during Virus Reconstitution

摘要: Recently the mouse cytomegalovirus (MCMV) genome was cloned as an infectious bacterial artificial chromosome (BAC) (M. Messerle, I. Crnkovic ´, W. Hammerschmidt, H. Ziegler, and U. Koszinowski, Proc. Natl. Acad. Sci. USA 94:14759‐14763, 1997). The virus obtained from this construct is attenuated in vivo due to deletion of viral sequences insertion BAC vector. We reconstituted full-length MCMV flanked vector with identical sequences. This new represents a versatile basis for construction mutants since generated loses acquires wild-type properties. Human (HCMV) ubiquitous pathogen that can cause severe disease immunologically immature immunocompromised patients (5). strict species specificity HCMV precludes investigation infection animal host. Infection valuable model studying various aspects CMV pathogenesis (13, 14). Fast efficient mutagenesis procedures are desirable order analyze role genes during course vivo. However, current schemes difficult, laborious, time-consuming replicates rather slowly relies on rare recombination events eukaryotic cells (12, 15, 18, 27, 28). Recently, we reported approach generation herpesvirus based homologous Escherichia coli (21). To end,