RECOMBITANT PLASMID DNA pET21-VP40VE CONTAINING THE GENE FOR THE MATRIX PROTEIN VP40 OF EBOLA VIRUS AND THE RECOMBITANT PROTEIN VP40-VE OBTAINED AS A RESULT OF THE GENE EXPRESSION FOR THE PROTEIN VP40 OF EBOLA VIRUS USING THE RECOMBITANT PLASMID DNA pET21-VP40VE AND EXHIBITING IMMUNOGENIC AND ANTIGENIC PROPERTIES Russian patent published in 2021 - IPC C12N15/70 

FIELD: biotechnology, medicine and molecular biology.

SUBSTANCE: invention relates in particular to the in vitro construction of a DNA construct pET21-VP40VE containing the gene for the matrix protein VP40 of the Ebola virus (strain Zaire, isolate of Mayinga) and recombinant protein VP40-VE, which has immunogenic and antigenic properties. The recombinant plasmid DNA pET21-VP40VE is intended for the expression of the Ebola virus matrix protein VP40 gene (strain Zaire, Mayinga isolate) in the E.coli prokaryotic system and for the production of the recombinant VP40-VE protein, which has a size of 6366 bp. and contains: the bla gene (coordinates 599 to 1459 bp) as a genetic marker that determines ampicillin resistance of E.coli (BL21/DE3(+) cells transformed with the recombinant plasmid pET21-VP40VE; site of origin replication ori (coordinates from 1630 to 2218 bp); phage T7 promoter (coordinates from 5117 to 5135 bp); lactose repressor (coordinates from 3648 bp to 4730 bp); DNA-binding protein that inhibits the expression of E.coli genes and ensures the expression of the target recombinant protein VP40-VE; unique recognition sites by restriction endonucleases with the following coordinates: NheI (5204 bp) and XhoI (6205 bp); VP40 VE gene (strain Zaire, Mayinga isolate) (981 bp), inserted into the expression cassette at the restriction sites NheI (5204 bp) and XhoI (6205 bp) (from 4479 to 5459 nucleotides of genomic RNA), which encodes 327 amino acid residues (a.a.) of the viral matrix protein VP40; sequence (18 bp) encoding polyhistidine d (6xHis) polytract of six histidines at the N-end of the encoded protein for its subsequent purification using the method of Ni affinity chromatography. The recombinant VP40-VE protein was obtained as a result of the expression of the Ebola virus matrix protein VP40 gene (strain Zaire, Mayinga isolate) in the bacterial expression system of E.coli as part of the recombinant DNA construct pET21-VP40VE, having a molecular weight of 36114.68 Da and the amino acid sequence SEQ ID NO: 1 333 a.a. length, including 6 histidines and a stop codon, and has immunogenic and antigenic properties and is intended for the induction of antibodies in the body of Balb/c mice that recognize the viral protein VP40-BE in immunochemical reactions, as well as interacts with mouse monoclonal antibodies, polyclonal goat and human antibodies specific for inactivated and infectious Ebola virus.

EFFECT: increased yield of target recombinant matrix protein VP40 VE.

2 cl, 14 dwg, 3 tbl, 8 ex