METHOD FOR PRODUCING RECOMBINANT HERPESVIRUSES FOR SIMULTANEOUS TRANSDUCTION AND IMMORTALIZATION OF PRIMARY T-LYMPHOCYTES AND NK CELLS Russian patent published in 2021 - IPC C12N15/64 C12N15/39 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, namely to a method for producing recombinant herpesviruses for simultaneous transduction and immortalization of primary T-lymphocytes and NK cells. The recombinant plasmid DNA of pUC-HVS-OFP consists of the following key genetic elements: puromycin resistance gene (pac); orange fluorescent protein (OFP) gene; flanking parts (600 bp) homologous to vCD59 area of HVS genome; ampicillin antibiotic resistance gene (AmpR) and bacterial promoter of ampicillin resistance gene (AmpR promoter); origin of replication of bacteriophage f1 (ori); cytomegalovirus (CMV) promoter. A cell culture permissive to lytic infection with Herpesvirus Saimiri virus is infected with Herpesvirus Saimiri HVS-C488 virus strain. Another interweaved cell line is transfected with a mixture of expression plasmids and pUC-HVS-OFP plasmid. Then the permissive cell culture infected with Herpesvirus Saimiri and the cell culture transfected with expression plasmids are mixed and subjected to co-cultivation.

EFFECT: combining the productive development of the virus in the cells of lines permissive to lytic HVS infection with the high efficiency of cell transfection with expression plasmids encoding genetic sequences intended for introduction into genome of the “wild” virus strain makes it possible to obtain recombinant herpesviruses with increased efficiency of development.

6 cl, 6 dwg