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Cited by 317

The DOI matches its Crossref record; 25 of the 25 references it cites were found in a registry or an index.

Cloning and Mutagenesis of the Murine Gammaherpesvirus 68 Genome as an Infectious Bacterial Artificial Chromosome

Open access
Adler, Heiko, Messerle, Martin, Wagner, Markus, Koszinowski, Ulrich H.·2000·Journal of Virology

Abstract

Gammaherpesviruses cause important infections of humans, in particular in immunocompromised patients. Recently, murine gammaherpesvirus 68 (MHV-68) infection of mice has been developed as a small animal model of gammaherpesvirus pathogenesis. Efficient generation of mutants of MHV-68 would significantly contribute to the understanding of viral gene functions in virus-host interaction, thereby further enhancing the potential of this model. To this end, we cloned the MHV-68 genome as a bacterial artificial chromosome (BAC) in Escherichia coli . During propagation in E. coli , spontaneous recombination events within the internal and terminal repeats of the cloned MHV-68 genome, affecting the copy number of the repeats, were occasionally observed. The gene for the green fluorescent protein was incorporated into the cloned BAC for identification of infected cells. BAC vector sequences were flanked by loxP sites to allow the excision of these sequences using recombinase Cre and to allow the generation of recombinant viruses with wild-type genome properties. Infectious virus was reconstituted from the BAC-cloned MHV-68. Growth of the BAC-derived virus in cell culture was indistinguishable from that of wild-type MHV-68. To assess the feasibility of mutagenesis of the cloned MHV-68 genome, a mutant virus with a deletion of open reading frame 4 was generated. Genetically modified MHV-68 can now be analyzed in functionally modified mouse strains to assess the role of gammaherpesvirus genes in virus-host interaction and pathogenesis.

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Crossref · 10.1128/jvi.74.15.6964-6974.2000

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Metadata

Authors
Adler, H., Messerle, M., Wagner, M., & Koszinowski, U. H.
Venue
Journal of Virology
Published
2000-08
Volume
74
Issue
15
Pages
6964-6974
ISSN
  • 0022-538X (ISSN-L)
  • 1098-5514
OpenAlex
W2138842966
Cited by
317
References
25
FWCI
4.59
Type
journal article
Language
en
Open access
green

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CoChat. (2026). Citation check: Cloning and Mutagenesis of the Murine Gammaherpesvirus 68 Genome as an Infectious Bacterial Artificial Chromosome. https://app.cochat.ai/papers/10.1128/jvi.74.15.6964-6974.2000
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Adler, H., Messerle, M., Wagner, M., & Koszinowski, U. H. (2000). Cloning and Mutagenesis of the Murine Gammaherpesvirus 68 Genome as an Infectious Bacterial Artificial Chromosome. Journal of Virology. https://doi.org/10.1128/jvi.74.15.6964-6974.2000

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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.