Fachbereich Veterinärmedizin



    Cloning the simian varicella virus genome in E. coli as an infectious bacterial artificial chromosome (2011)

    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Gray, Wayne L
    Zhou, Fuchun
    Noffke, Juliane
    Tischer, B Karsten
    Archives of virology; 156(5) — S. 739–746
    ISSN: 0304-8608
    DOI: 10.1007/s00705-010-0889-4
    Pubmed: 21487663
    Institut für Virologie

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    Abstract / Zusammenfassung

    Simian varicella virus (SVV) is closely related to human varicella-zoster virus and causes varicella and zoster-like disease in nonhuman primates. In this study, a mini-F replicon was inserted into a SVV cosmid, and infectious SVV was generated by co-transfection of Vero cells with overlapping SVV cosmids. The entire SVV genome, cloned as a bacterial artificial chromosome (BAC), was stably propagated upon serial passage in E. coli. Transfection of pSVV-BAC DNA into Vero cells yielded infectious SVV (rSVV-BAC). The mini-F vector sequences flanked by loxP sites were removed by co-infection of Vero cells with rSVV-BAC and adenovirus expressing Cre-recombinase. Recombinant SVV generated using the SVV-BAC genetic system has similar molecular and in vitro replication properties as wild-type SVV. To demonstrate the utility of this approach, a SVV ORF 10 deletion mutant was created using two-step Red-mediated recombination. The results indicate that SVV ORF 10, which encodes a homolog of the HSV-1 virion VP-16 transactivator protein, is not essential for in vitro replication but is required for optimal replication in cell culture.