Journal
JOURNAL OF BIOMEDICINE AND BIOTECHNOLOGY
Volume -, Issue -, Pages -Publisher
HINDAWI PUBLISHING CORPORATION
DOI: 10.1155/2011/926258
Keywords
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Funding
- Leukemia and Lymphoma Society
- NIH [R21 DE018337, DE015752, DE19085]
- STOP Cancer Foundation
- Burroughs Wellcome Fund
- NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH [P01DE019085] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF DENTAL &CRANIOFACIAL RESEARCH [R01DE015752, R21DE018337] Funding Source: NIH RePORTER
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Here we describe the cloning of a sequenced WUMS isolate of murine gammaherpesvirus-68 (MHV-68, gamma HV-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.
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