SET OF RECOMBINANT PLASMID DNA FOR OBTAINING RECOMBINANT VIRUSES SENDAI STRAIN MOSCOW (OPTIONS) Russian patent published in 2023 - IPC C12N7/01 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. A set of recombinant plasmid DNA for obtaining recombinant Sendai viruses of the Moscow strain is presented. The invention relates to recombinant plasmid DNA containing a full-length DNA copy of the genome of the Sendai virus strain Moscow and genes for the structural proteins of nucleocapsid N, phosphoprotein P and viral polymerase L, and can be used in biotechnology, in particular in genetic engineering for constructing recombinant variants of the Sendai virus strain Moscow, containing inserts of transgenes of antitumor proteins or protectively significant antigens of pathogenic agents, in order to develop antitumor drugs and vector vaccines against infectious diseases on their basis. The structural element includes a polylinker of five unique sites for large-cleaved restriction endonucleases BsiWI, NruI, ClaI, AscI, BssHII, followed by a transgene transcription termination signal and a subsequent gene transcription reinitiation signal for Sendai virus RNA polymerase. The expression of the N, P, and L genes of the Sendai virus of the Moscow strain, as well as the synthesis of genomic RNA of recombinant strains of the Sendai virus, is carried out from the corresponding plasmid DNA under the control of T7 phage polymerase. The size accuracy of genomic RNA is controlled by ribozymes inserted at its 3' and 5' ends. The technical result of the invention is the production of 3 recombinant plasmid DNA expressing the N, P and L genes of the Sendai virus of the Moscow strain (auxiliary plasmids pET15b-N, pET15b-P, pET15b-L), as well as two plasmids with full-length copies of DNA of the genomic RNA of the virus Sendai strain Moscow (GenBank Asc. KP717417.1) containing a structural element for the insertion and expression of transgenes in the 3'-untranslated region of the viral genome (plasmid pSen2-MSC(N)) and between the P and M genes (pSen2-MCS(M)). A set of three auxiliary plasmids and a genomic plasmid pSen2-MSC(N) provides the creation of recombinant variants of the Sendai virus of the Moscow strain with a high level of transgene expression. A set of the same auxiliary plasmids and the pSen2-MSC(M) genomic plasmid ensures the creation of recombinant variants of the Sendai virus of the Moscow strain with an average level of transgene expression. Verification of the proposed set of recombinant plasmid DNA was carried out on the example of a model transgene of the green fluorescent protein EGFP.

EFFECT: creation of set of recombinant plasmid DNA for obtaining recombinant Sendai viruses of the Moscow strain.

2 cl, 9 dwg, 3 tbl, 3 ex