Reverse genetic operating system for segmented cloning and rescue of pseudorabies virus BarthaK61 vaccine strain genome as well as construction method and application of reverse genetic operating system

The invention relates to a reverse genetic operating system for segmented cloning and rescue of a pseudorabies virus BarthaK61 vaccine strain genome as well as a construction method and application of the reverse genetic operating system. The system comprises recombinant plasmids BAC-cm-PRV-EA, BAC-cm-PRV-BC and BAC-cm-PRV-FG which contain the whole genome of the pseudorabies virus Barra K61 vaccine strain, and a host cell for expressing the whole genome of the pseudorabies virus Barra K61 vaccine strain. According to the reverse genetic manipulation system of the pseudorabies virus BarthaK61 vaccine strain, a BarthaK61 genome is successfully split into six segments through an ExoCET cloning and assembling technology, then the six segments are assembled into three recombinant plasmids BAC-cm-PRV-EA, BAC-cm-PRV-BC and BAC-cm-PRV-FG, the three recombinant plasmids can be jointly transfected into African green monkey kidney cells after being linearized through PacI, the BarthaK61 virus is rescued, and the recomb