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Recombinant Modified Vaccinia Virus Ankara Development to Express VP2, NS1, and VP7 Proteins of Bluetongue Virus

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Vaccine Technologies for Veterinary Viral Diseases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2465))

Abstract

Modified vaccinia virus Ankara (MVA) is employed widely as an experimental vaccine vector for its abortive replication in mammalian cells and high expression level of foreign/heterologous genes. Recombinant MVAs (rMVAs) are used as platforms for protein production as well as vectors to generate vaccines against a wide range of infectious diseases and other pathologies. The portrait of the virus combines desirable elements such as high-level biological safety, the ability to activate appropriate innate immune mediators upon vaccination , and the capacity to deliver substantial amounts of heterologous antigens. rMVAs encoding proteins of Bluetongue virus (BTV), an orbivirus that infects domestic and wild ruminants through transmission by biting midges of the Culicoides species, are excellent vaccine candidates against this virus. In this chapter, we describe the methods for the generation of rMVAs encoding VP2, NS1, and VP7 proteins of BTV . The included protocols cover the cloning of VP2, NS1, and VP7 BTV-4 genes in a transfer plasmid, the construction of rMVAs, the titration of virus working stocks, and the protein expression analysis by immunofluorescence and radiolabeling of rMVA infected cells as well as virus purification procedure.

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Authors and Affiliations

  1. Section of Infectious Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT, USA

    Alejandro Marín-López

  2. Centro de Investigación en Sanidad Animal (CISA), INIA-CSIC, Madrid, Spain

    Sergio Utrilla-Trigo, Luis Jiménez-Cabello & Javier Ortego

Authors
  1. Alejandro Marín-López
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  2. Sergio Utrilla-Trigo
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  3. Luis Jiménez-Cabello
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  4. Javier Ortego
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Corresponding author

Correspondence to Javier Ortego .

Editor information

Editors and Affiliations

  1. Centro de Investigación en Sanidad Animal (CISA), INIA-CSIC, Valdeolmos, Madrid, Spain

    Alejandro Brun

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Marín-López, A., Utrilla-Trigo, S., Jiménez-Cabello, L., Ortego, J. (2022). Recombinant Modified Vaccinia Virus Ankara Development to Express VP2, NS1, and VP7 Proteins of Bluetongue Virus. In: Brun, A. (eds) Vaccine Technologies for Veterinary Viral Diseases. Methods in Molecular Biology, vol 2465. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2168-4_10

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  • DOI: https://doi.org/10.1007/978-1-0716-2168-4_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2167-7

  • Online ISBN: 978-1-0716-2168-4

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