The Pirbright Institute, Pirbright, Woking, UK.
Methods Mol Biol. 2022;2503:73-94. doi: 10.1007/978-1-0716-2333-6_5.
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).
ASFV 的基因操作不仅越来越多地用于开发活减毒疫苗,而且还作为进一步了解病毒-宿主相互作用的不可或缺的工具。在这里,我们介绍了使用显色和荧光报告基因分离猪骨髓细胞和纯化重组 ASFV 的方法。我们还详细描述了一种新开发的使用荧光激活细胞分选(FACS)纯化遗传修饰的 ASFV 的方法。