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

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. I...

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Detalles Bibliográficos
Autores principales: Labroussaa, Fabien, Mehinagic, Kemal, Cippa, Valentina, Liniger, Matthias, Akarsu, Hatice, Ruggli, Nicolas, Jores, Joerg
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8433286/
https://www.ncbi.nlm.nih.gov/pubmed/34527959
http://dx.doi.org/10.1016/j.xpro.2021.100803
Descripción
Sumario: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.

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