Construction and characterization of an infectious murine gammaherpesivrus-68 bacterial artificial chromosome

Ting Ting Wu*, Hsiang I. Liao, Leming Tong, Ronika Sitapara Leang, Greg Smith, Ren Sun

*Corresponding author for this work

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Here we describe the cloning of a sequenced WUMS isolate of murine gammaherpesvirus-68 (MHV-68, γHV-68, also known as MuHV-4) as a bacterial artificial chromosome (BAC). We engineered the insertion of the BAC sequence flanked by loxP sites into the left end of the viral genome before the M1 open reading frame. The infectious viruses were reconstituted following transfection of the MHV-68 BAC DNA into cells. The MHV-68 BAC-derived virus replicated indistinguishably from the wild-type virus in cultured cells. Excision of the BAC insert was efficiently achieved by coexpressing the Cre recombinase. Although the BAC insertion did not significantly affect acute productive infection in the lung, it severely compromised the ability of MHV-68 to establish splenic latency. Removal of the BAC sequence restored the wild-type level of latency. Site-specific mutagenesis was carried out by RecA-mediated recombination to demonstrate that this infectious BAC clone can be used for genetic studies of MHV-68.

Original languageEnglish (US)
Article number926258
JournalJournal of Biomedicine and Biotechnology
Volume2011
DOIs
StatePublished - 2011

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Health, Toxicology and Mutagenesis

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