Viruses, Vol. 12, Pages 615: Production of Recombinant African Swine Fever Viruses: Speeding Up the Process
Viruses, Vol. 12, Pages 615: Production of Recombinant African Swine Fever Viruses: Speeding Up the Process
Viruses doi: 10.3390/v12060615
Authors:
Anusyah Rathakrishnan
Katy Moffat
Ana Luisa Reis
Linda K. Dixon
African swine fever (ASF) is a devastating disease in pigs, with no vaccines for control. The genetic manipulation of African swine fever virus (ASFV) is often tedious and time consuming. Here, we describe a method to manipulate the virus genome to produce gene deletion viruses in a much-reduced time. This method combines the conventional homologous recombination with fluorescent-activated cells sorting (FACS), to isolate and purify viruses expressing fluorescent reporter genes. With three rounds of single cell isolation via FACS and two rounds of limiting dilution, we deleted two additional genes, EP153R and EP402R, from Benin 97/1 ASFV lacking the DP148R gene. By combining different fluorescent markers, this method has the potential to greatly facilitate studies on understanding ASFV gene functions and develop candidate live-attenuated vaccines.
Source: Viruses - Category: Virology Authors: Anusyah Rathakrishnan Katy Moffat Ana Luisa Reis Linda K. Dixon Tags: Article Source Type: research