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Construction and Characterization of Bacterial Artificial Chromosomes (BACs) Containing Herpes Simplex Virus Full-Length Genomes

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Herpes Simplex Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1144))

Abstract

Bacterial artificial chromosomes (BACs) are suitable vectors not only to maintain the large genomes of herpesviruses in Escherichia coli but also to enable the traceless introduction of any mutation using modern tools of bacterial genetics. To clone a herpes simplex virus genome, a BAC replication origin is first introduced into the viral genome by homologous recombination in eukaryotic host cells. As part of their nuclear replication cycle, genomes of herpesviruses circularize and these replication intermediates are then used to transform bacteria. After cloning, the integrity of the recombinant viral genomes is confirmed by restriction length polymorphism analysis and sequencing. The BACs may then be used to design virus mutants. Upon transfection into eukaryotic cells new herpesvirus strains harboring the desired mutations can be recovered and used for experiments in cultured cells as well as in animal infection models.

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Author information

Authors and Affiliations

  1. Heinrich Pette Institute—Leibniz-Institute for Experimental Virology, Hamburg, Germany

    Claus-Henning Nagel

  2. Institute of Virology, OE 5230, Hannover Medical School, Carl-Neuberg-Str. 1, Hannover, 30625, Germany

    Anja Pohlmann & Beate Sodeik

Authors
  1. Claus-Henning Nagel
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  2. Anja Pohlmann
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  3. Beate Sodeik
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Corresponding author

Correspondence to Beate Sodeik .

Editor information

Editors and Affiliations

  1. Centre for Virus Research, Westmead Millennium Institute, Westmead, New South Wales, Australia

    Russell J. Diefenbach

  2. Institute of Virology, University of Zürich, Zürich, Switzerland

    Cornel Fraefel

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Nagel, CH., Pohlmann, A., Sodeik, B. (2014). Construction and Characterization of Bacterial Artificial Chromosomes (BACs) Containing Herpes Simplex Virus Full-Length Genomes. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus. Methods in Molecular Biology, vol 1144. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0428-0_4

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  • DOI: https://doi.org/10.1007/978-1-4939-0428-0_4

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0427-3

  • Online ISBN: 978-1-4939-0428-0

  • eBook Packages: Springer Protocols

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