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Coronaviruses pp 167–184Cite as

In-Yeast Assembly of Coronavirus Infectious cDNA Clones Using a Synthetic Genomics Pipeline

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2203))

Abstract

The Escherichia coli and vaccinia virus-based reverse genetics systems have been widely applied for the manipulation and engineering of coronavirus genomes. These systems, however, present several limitations and are sometimes difficult to establish in a timely manner for (re-)emerging viruses. In this chapter, we present a new universal reverse genetics platform for the assembly and engineering of infectious full-length cDNAs using yeast-based transformation-associated recombination cloning. This novel assembly method not only results in stable coronavirus infectious full-length cDNAs cloned in the yeast Saccharomyces cerevisiae but also fosters and accelerates the manipulation of their genomes. Such a platform is widely applicable for the scientific community, as it requires no specific equipment and can be performed in a standard laboratory setting. The protocol described can be easily adapted to virtually all known or emerging coronaviruses, such as Middle East respiratory syndrome coronavirus (MERS-CoV).

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Author information

Author notes

  1. Tran Thi Nhu Thao, Fabien Labroussaa, Joerg Jores, and Volker Thiel contributed equally to this work.

Authors and Affiliations

  1. Institute of Virology and Immunology IVI, Bern, Switzerland

    Tran Thi Nhu Thao, Nadine Ebert & Volker Thiel

  2. Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland

    Tran Thi Nhu Thao, Fabien Labroussaa, Nadine Ebert, Joerg Jores & Volker Thiel

  3. Graduate School for Biomedical Science, University of Bern, Bern, Switzerland

    Tran Thi Nhu Thao

  4. Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Bern, Bern, Switzerland

    Fabien Labroussaa & Joerg Jores

Authors
  1. Tran Thi Nhu Thao
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  2. Fabien Labroussaa
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  3. Nadine Ebert
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  4. Joerg Jores
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  5. Volker Thiel
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Corresponding authors

Correspondence to Joerg Jores or Volker Thiel .

Editor information

Editors and Affiliations

  1. The Pirbright Institute, Surrey, UK

    Helena J. Maier

  2. The Pirbright Institute, Surrey, UK

    Erica Bickerton

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Thao, T.T.N., Labroussaa, F., Ebert, N., Jores, J., Thiel, V. (2020). In-Yeast Assembly of Coronavirus Infectious cDNA Clones Using a Synthetic Genomics Pipeline. In: Maier, H., Bickerton, E. (eds) Coronaviruses. Methods in Molecular Biology, vol 2203. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0900-2_13

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  • DOI: https://doi.org/10.1007/978-1-0716-0900-2_13

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0899-9

  • Online ISBN: 978-1-0716-0900-2

  • eBook Packages: Springer Protocols

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