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

STAR Protoc. 2021-09;

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

Products/Services Used	Details	Operation
Recombinant Proteins	… used in this protocol involves the fragmentation of the ASFV genome into seven sub-genomic fragments as presented in Figure 1. 1. The 5′ and 3′-ends of the ASFV genome, consisting of inverted-terminal-repeats (ITRs), are chemically-synthetized (GenScript) in order to …	Get A Quote

摘要

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.