ARTIFICIAL GENE CODING A BICISTRONIC STRUCTURE FORMED BY RECEPTOR-BINDING DOMAIN SEQUENCES OF THE GLYCOPROTEIN S OF THE SARS-COV-2 CORONAVIRUS, P2A-PEPTIDE AND GLYCOPROTEIN G VSV, RECOMBINANT PLASMID PSTEM-RVSV-STBL_RBD_SC2, PROVIDING EXPRESSION OF ARTIFICIAL GENE, AND A RECOMBINANT STRAIN OF VESICULAR STOMATITIS VIRUS RVSV-STBL_RBD_SC2, USED TO CREATE A VACCINE AGAINST SARS-COV-2 CORONAVIRUS
Imatdinov Ilnaz Ramisovich (RU),Иматдинов Ильназ Рамисович (RU),Bochkareva Mariya Dmitrievna (RU),Бочкарева Мария Дмитриевна (RU),Prudnikova Elena Yurevna (RU),Прудникова Елена Юрьевна (RU),Tishin Ant
申请号:
RU2020123453
公开号:
RU0002733831C1
申请日:
2020.07.08
申请国别(地区):
RU
年份:
2020
代理人:
摘要:
FIELD: biotechnology; molecular biology; genetic engineering; medicine.SUBSTANCE: described is an artificial gene used to create a vaccine against SARS-CoV-2 coronavirus, coding an artificial protein-immunogen, which is a bicistronic structure consisting of receptor-binding domain (RBD) sequences of SARS-CoV-2 coronavirus glycoprotein S, heterological signal peptide haemagglutinin (HA) of influenza A virus, a linker, P2A-peptide for splitting polyprotein during translation and glycoprotein G with mutation M(1)>;P(1), preventing alternative translation initiation, presented in SEQ ID NO: 1, length of 2355 bp. Disclosed is a recombinant plasmid pStem-rVSV-Stbl_RBD_SC2, used to create a vaccine against SARS-CoV-2 coronavirus, having molecular weight 8.85⋅106 Da, size 14309 bp and comprising, in accordance with physical and genetic map presented in Fig. 2, target gene according to item 1, coding artificial protein-immunogen RBD of SARS-CoV-2 coronavirus glycoprotein S, having amino acid sequence SEQ ID NO: 2 and under control of viral promoter providing its expression in mammalian cells. Strain rVSV-Stbl_RBD_SC2 of a recombinant virus of vesicular stomatitis obtained using a recombinant plasmid pStem-rVSV-Stbl_RBD_SC2, providing independent synthesis of coronavirus antigen (receptor-binding domain of SARS-CoV-2 glycoprotein S) and protein G of virus of vesicular stomatitis used to develop a vaccine against SARS-CoV-2 coronavirus and deposited in the State collection of viral infection causative agents, rickettsioses of State Research Center of Virology and Biotechnology VECTOR of Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, No. V-984.EFFECT: improved folding and higher infectious titre of the recombinant virus, stability of the target transgene and immunogenic / antigenic properties of the declared recombinant virus.3 cl, 5 dwg, 3 tbl, 5 exИзобретение относится к биотехнологии, молекулярной биологии, генетической инженерии и меди