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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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
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author Labroussaa, Fabien
Mehinagic, Kemal
Cippa, Valentina
Liniger, Matthias
Akarsu, Hatice
Ruggli, Nicolas
Jores, Joerg
author_facet Labroussaa, Fabien
Mehinagic, Kemal
Cippa, Valentina
Liniger, Matthias
Akarsu, Hatice
Ruggli, Nicolas
Jores, Joerg
author_sort Labroussaa, Fabien
collection PubMed
description 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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spelling pubmed-84332862021-09-14 In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples Labroussaa, Fabien Mehinagic, Kemal Cippa, Valentina Liniger, Matthias Akarsu, Hatice Ruggli, Nicolas Jores, Joerg STAR Protoc Protocol 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. Elsevier 2021-09-08 /pmc/articles/PMC8433286/ /pubmed/34527959 http://dx.doi.org/10.1016/j.xpro.2021.100803 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Labroussaa, Fabien
Mehinagic, Kemal
Cippa, Valentina
Liniger, Matthias
Akarsu, Hatice
Ruggli, Nicolas
Jores, Joerg
In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples
title In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples
title_full In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples
title_fullStr In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples
title_full_unstemmed In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples
title_short In-yeast reconstruction of the African swine fever virus genome isolated from clinical samples
title_sort in-yeast reconstruction of the african swine fever virus genome isolated from clinical samples
topic Protocol
url 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
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