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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
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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.

关键词

Biotechnology and bioengineering, Genomics, Health Sciences, Microbiology, Molecular Biology, Sequencing, Systems biology
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