title: Cloning the simian varicella virus genome in E. coli as an infectious bacterial artificial chromosome.
abstract: 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.
authors.affiliation: Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, 72205, USA. graywaynel@uams.edu
authors.forename: Wayne L
authors.initials: WL
authors.lastname: Gray
authors.forename: Fuchun
authors.initials: F
authors.lastname: Zhou
authors.forename: Juliane
authors.initials: J
authors.lastname: Noffke
authors.forename: B Karsten
authors.initials: BK
authors.lastname: Tischer
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D022202:Chromosomes, Artificial, Bacterial; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D016679:Genome, Viral; D006564:Herpesviridae / Q000235:genetics*; D014709:Vero Cells