Labroussaa, Fabien; Mehinagic, Kemal; Cippa, Valentina; Liniger, Matthias; Akarsu, Hatice; Ruggli, Nicolas; Jores, Joerg (2021). In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples. STAR protocols, 2(3), p. 100803. Elsevier 10.1016/j.xpro.2021.100803
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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.