A synthetic genomics pipeline using transformation-associated recombination cloning successfully cloned and engineered the entire 190-kbp genome of the African swine fever virus genotype II in yeast.
This protocol enables the cloning and engineering of large DNA viruses like ASFV in yeast, facilitating viral genomics research.
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.
Labroussaa et al. (2021) studied African swine fever virus (ASFV). Synthetic genomics pipeline (transformation-associated recombination cloning) was evaluated on Cloning and engineering of the entire 190-kbp genome of ASFV genotype II in yeast. A synthetic genomics pipeline using transformation-associated recombination cloning successfully cloned and engineered the entire 190-kbp genome of the African swine fever virus genotype II in yeast.
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