FIELD: biotechnology.
SUBSTANCE: invention relates to the field of biotechnology. Invention is a recombinant plasmid DNA pET21-NPVE for expression of the nucleoprotein gene NP of Ebola virus (Zaire strain, Mayinga isolate) in a prokaryotic system of E.coli and producing recombinant protein NP-VE, has size of 7593 base pairs and contains: bla gene (coordinates from 599 to 1459 base pairs) as a genetic marker; a replication starting point ori (coordinates from 1630 to 2218 base pairs); phage T7 promoter (coordinates from 5117 to 5135 base pairs; lactose repressor (coordinates with 3648 base pairs at 4730 base pairs); DNA-binding protein which inhibits expression of E.coli genes and provides expression of the target recombinant NP-VE protein after addition of an isopropyl-β-D-1-thiogalactopyranoside (IPTG); unique recognition sites with restriction endonucleases, having the following coordinates: NheI (5209 base pair) and Xho I (7432 base pairs); full-length NP gene of Ebola virus (Mayinga isolate, Zaire strain) 2217 base pairs (from 470 to 2686 nucleotide genomic RNA), built into the expression cassette by NheI restriction sites (5209 base pair) and XhoI (7432 base pairs), which encodes 739 amino acid residues (ao) of viral nucleoprotein; sequence (18 base pairs), encoding polyhistidine (6xHis) polithract of six histidines on the N-end of the encoded protein for subsequent purification by affinity N-chromatography and a recombinant NP-VE protein produced as a result of Ebola virus nucleoprotein NP gene expression (Zaire strain, Mayinga isolate) in the bacterial expression system E.coli in the recombinant DNA construct pET21-NPVE having a calculated molecular weight of 84457.42 Da, and amino acid sequence SEQ ID NO: 1 length of 748 amino-acid residues, including 6xHis, and a stop codon, and having immunogenic and antigenic properties, and intended for induction in the body of Balb/c mice of antibodies recognizing in immunochemical reactions viral protein NP-VE, as well as interacting with mouse monoclonal antibodies, polyclonal goat and human antibodies specific to inactivated and infectious virus, respectively.
EFFECT: invention increases output of the target recombinant protein NP-VE.
2 cl, 3 tbl, 14 dwg
| Title | Year | Author | Number |
|---|---|---|---|
| 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 | 2020 |
|
RU2742511C1 |
| STRAIN OF HYBRID ANIMAL CELLS Mus musculus L - PRODUCER OF MONOCLONAL ANTIBODIES FOR EXPOSING VP40 PROTEIN OF EBOLA VIRUS, ZAIRE SUBTYPE (Mainga STRAIN) (VERSIONS), MONOCLONAL ANTIBODY PRODUCED BY SAID STRAIN (VERSIONS) AND SET FOR "SANDWICH" FORMAT IMMUNOENZYMOMETRIC TEST SYSTEM FOR EXPOSING VP40 PROTEIN OF EBOLA VIRUS, ZAIRE SUBTYPE (Mainga STRAIN) | 2008 |
|
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|
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| STRAIN OF HYBRID ANIMAL CELLS Mus museums L - PRODUCER OF MONOCLONAL ANTIBODIES FOR EXPOSING NUCLEOPROTEIN OF EBOLA VIRUS, ZAIRE SUBTYPE (Mainga STRAIN) (VERSIONS), MONOCLONAL ANTIBODY PRODUCED BY SAID STRAIN (VERSIONS) AND SET FOR "SANDWICH" FORMAT IMMUNOENZYMOMETRIC TEST SYSTEM FOR EXPOSING NUCLEOPROTEIN OF EBOLA VIRUS, ZAIRE SUBTYPE (Mainga STRAIN) | 2008 |
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RU2395576C1 |
| HSOBIENS-WT/GIN/2015/KALIDIE-KINDIA-1022 STRAIN OF EBOLA ZAIRE TO OBTAIN ANTIGEN USED AS COMPONENT OF IMMUNOENZYMOMETRIC TEST SYSTEM FOR DETECTION OF G AND M ANTIBODIES TO VIRUS OF EBOLA | 2016 |
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| SINGLE-DOMAIN ANTIBODIES TO EBOLA VIRUS GP PROTEIN FOR IMMUNOTHERAPY OF EBOLA FEVER | 2015 |
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| pET21-GST-CD RECOMBINANT PLASMID WHICH ENSURES SYNTHESIS AND SECRETION OF Gn ECTODOMAIN OF HANTAAN VIRUS GLYCOPROTEIN, AND BL21(DE3)PLYSE pET21-GST-CD RECOMBINANT ESCHERICHIA COLI STRAIN, PRODUCER OF Gn PROTEIN, ECTODOMAIN OF HANTAAN VIRUS GLYCOPROTEIN | 2023 |
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| RECOMBINANT PLASMID pET21-GST-CD WHICH ENSURES THE SYNTHESIS AND SECRETION OF Gc ECTODOMAIN OF HANTAAN VIRUS GLYCOPROTEIN, AND THE RECOMBINANT ESCHERICHIA COLI STRAIN BL21(DE3)pLysE pET21-GST-CD, A PRODUCER OF THE Gc PROTEIN, THE ECTODOMAIN OF HANTAAN VIRUS GLYCOPROTEIN | 2023 |
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RU2807520C1 |
| ARTIFICIAL GENES CODING EV.CTL AND EV.Th PROTEINS-IMMUNOGENS, RECOMBINANT PLASMID DNA pEV.CTL AND EV.Th, PROVIDING EXPRESSION OF ARTIFICIAL GENES, AND ARTIFICIAL T-CELL POLYEPITOPE PROTEINS-IMMUNOGENS EV.CTL AND EV.Th CONTAINING EPITOPES OF EBOLA VIRUS ANTIGENS, USED TO CREATE AN EBOLA VIRUS VACCINE | 2018 |
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| STRAIN OF EBOL VIRUS "ZAIR K-5" FOR CARRYING OUT MODEL EXPERIMENTS AND PREPARING DIAGNOSTIC AND VACCINE PREPARATIONS | 2001 |
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