Isolation of Porcine Bone Marrow Cells and Generation of Recombinant African Swine Fever Viruses

Anusyah Rathakrishnan; Ana Luisa Reis; Katy Moffat; Linda K Dixon

doi:10.1007/978-1-0716-2333-6_5

Isolation of Porcine Bone Marrow Cells and Generation of Recombinant African Swine Fever Viruses

Methods Mol Biol. 2022:2503:73-94. doi: 10.1007/978-1-0716-2333-6_5.

Authors

Anusyah Rathakrishnan¹, Ana Luisa Reis², Katy Moffat², Linda K Dixon²

Affiliations

¹ The Pirbright Institute, Pirbright, Woking, UK. anusyah.rathakrishnan@pirbright.ac.uk.
² The Pirbright Institute, Pirbright, Woking, UK.

PMID: 35575887
DOI: 10.1007/978-1-0716-2333-6_5

Abstract

Genetic manipulation of ASFV has been increasingly used not only for the development of live attenuated vaccines but also as an indispensable tool to further our understanding of the virus-host interactions. Here we present methods for isolation of porcine bone marrow cells and purification of recombinant ASFV using both chromogenic and fluorescent reporters. We also describe in detail a newly developed method to purify genetically modified ASFV using fluorescence-activated cell sorting (FACS).

Keywords: ASFV; FACS; Porcine bone marrow cells; Recombinant virus; Single-cell isolation; Titration.

MeSH terms

African Swine Fever Virus* / genetics
African Swine Fever* / prevention & control
Animals
Bone Marrow Cells
Swine
Vaccines, Attenuated
Viral Proteins / genetics

Substances

Vaccines, Attenuated
Viral Proteins