In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples

Fabien Labroussaa; Kemal Mehinagic; Valentina Cippa; Matthias Liniger; Hatice Akarsu; Nicolas Ruggli; Joerg Jores

STAR Protocols (Sep 2021)

In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples

Fabien Labroussaa,
Kemal Mehinagic,
Valentina Cippa,
Matthias Liniger,
Hatice Akarsu,
Nicolas Ruggli,
Joerg Jores

Affiliations

Fabien Labroussaa: Institute of Veterinary Bacteriology, University of Bern, Bern 3001, Switzerland; Department of Infectious Diseases and Pathobiology (DIP), Vetsuisse Faculty, University of Bern, Bern 3001, Switzerland; Corresponding author
Kemal Mehinagic: Department of Infectious Diseases and Pathobiology (DIP), Vetsuisse Faculty, University of Bern, Bern 3001, Switzerland; Institute of Virology and Immunology (IVI), Sensemattstrasse 293, Mittelhäusern 3147, Switzerland; Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern 3001, Switzerland
Valentina Cippa: Institute of Veterinary Bacteriology, University of Bern, Bern 3001, Switzerland; Department of Infectious Diseases and Pathobiology (DIP), Vetsuisse Faculty, University of Bern, Bern 3001, Switzerland
Matthias Liniger: Department of Infectious Diseases and Pathobiology (DIP), Vetsuisse Faculty, University of Bern, Bern 3001, Switzerland; Institute of Virology and Immunology (IVI), Sensemattstrasse 293, Mittelhäusern 3147, Switzerland
Hatice Akarsu: Institute of Veterinary Bacteriology, University of Bern, Bern 3001, Switzerland; Department of Infectious Diseases and Pathobiology (DIP), Vetsuisse Faculty, University of Bern, Bern 3001, Switzerland
Nicolas Ruggli: Department of Infectious Diseases and Pathobiology (DIP), Vetsuisse Faculty, University of Bern, Bern 3001, Switzerland; Institute of Virology and Immunology (IVI), Sensemattstrasse 293, Mittelhäusern 3147, Switzerland
Joerg Jores: Institute of Veterinary Bacteriology, University of Bern, Bern 3001, Switzerland; Department of Infectious Diseases and Pathobiology (DIP), Vetsuisse Faculty, University of Bern, Bern 3001, Switzerland; Corresponding author

Journal volume & issue: Vol. 2, no. 3
p. 100803

Abstract

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Summary: This protocol describes a synthetic genomics pipeline to clone and engineer the entire 190-kbp genome of the African swine fever virus (ASFV) genotype II in yeast using transformation-associated recombination cloning. The viral genome was cloned using DNA directly extracted from a clinical sample. In addition, the precise deletion of a non-essential gene and its replacement by a synthetic reporter gene cassette are presented. This protocol is applicable to other ASFV genotypes and other large DNA viruses.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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