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

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

Summary: 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...

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Bibliographic Details
Main Authors: Fabien Labroussaa, Kemal Mehinagic, Valentina Cippa, Matthias Liniger, Hatice Akarsu, Nicolas Ruggli, Joerg Jores
Format: Article
Language:English
Published: Elsevier 2021-09-01
Series:STAR Protocols
Subjects:
Health Sciences
Genomics
Sequencing
Microbiology
Molecular Biology
Systems biology
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721005098
Description
Summary:Summary: 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.
ISSN:2666-1667

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