Publikationsdatenbank

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

Art

Zeitschriftenartikel / wissenschaftlicher Beitrag

Autoren

Gray, Wayne L

Zhou, Fuchun

Noffke, Juliane

Tischer, B Karsten

Quelle

Archives of virology : official journal of the Virology Division of the International Union of Microbiological Societies

Bandzählung: 156

Heftzählung: 5

Seiten: 739 – 746

ISSN: 0304-8608

Sprache

Englisch

Verweise

DOI: 10.1007/s00705-010-0889-4

Pubmed: 21487663

Kontakt

Institut für Virologie

Robert-von-Ostertag-Str. 7-13
14163 Berlin
+49 30 838 51833
virologie@vetmed.fu-berlin.de

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.