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

doi:10.1016/j.xpro.2021.100803

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

STAR Protoc. 2021 Sep 8;2(3):100803. doi: 10.1016/j.xpro.2021.100803. eCollection 2021 Sep 17.

Authors

Fabien Labroussaa^{1

2}, Kemal Mehinagic^{2

3

4}, Valentina Cippa^{1

2}, Matthias Liniger^{2

3}, Hatice Akarsu^{1

2}, Nicolas Ruggli^{2

3}, Joerg Jores^{1

2}

Affiliations

¹ Institute of Veterinary Bacteriology, University of Bern, Bern 3001, Switzerland.
² Department of Infectious Diseases and Pathobiology (DIP), Vetsuisse Faculty, University of Bern, Bern 3001, Switzerland.
³ Institute of Virology and Immunology (IVI), Sensemattstrasse 293, Mittelhäusern 3147, Switzerland.
⁴ Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern 3001, Switzerland.

Abstract

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.

Keywords: Biotechnology and bioengineering; Genomics; Health Sciences; Microbiology; Molecular Biology; Sequencing; Systems biology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

African Swine Fever / virology*
African Swine Fever Virus / genetics*
Animals
DNA, Viral / genetics
Genetic Engineering / methods*
Genomics
Saccharomyces cerevisiae / genetics*
Swine
Synthetic Biology

Substances

DNA, Viral