Author: Anusyah Rathakrishnan, Katy Moffat, Ana Luisa Reis and Linda K. Dixon
Year: 2020
About this Publication:
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.
Grant: VITAL
Subject Areas: Research and Development
Diseases: African Swine Fever
URL https://www.mdpi.com/1999-4915/12/6/615
Keywords:
ASFV, Benin 97/1 genotype I) BeninDDP148R, BeninDEP402R, DP148R, EP153R, EP402R, FACS, recombinant
Countries:
Benin, Botswana, Burkina Faso, Burundi, Cape Verde, Comoros, Cote D'Ivoire, Djibouti, Eritrea, Ethiopia, Gambia, Ghana, Guinea, Guinea-Bissau, Kazakhstan, Kenya, Kyrgyzstan, Lesotho, Liberia, Madagascar, Malawi, Mali, Mauritania, Mauritius, Mayotte, Mozambique, Namibia, Niger, Nigeria, Reunion, Rwanda, Saint Helena, Senegal, Seychelles, Sierra Leone, Somalia, South Africa, Swaziland, Tajikistan, Tanzania, United Republic Of, Togo, Turkmenistan, Uganda, Uzbekistan, Zambia, Zimbabwe
