FIELD: medicine.
SUBSTANCE: described is recombinant plasmid DNA pET-32a/TβRII. Plasmid consists of BgHI/Hindlll-fragment of plasmid pET-32a DNA, containing lacl-promoter E.coli, T7 promoter, Trx-Tag coding sequence, His-Tag coding sequence, S-Tag coding sequence, T7 terminator, lad-coding sequence, ba-coding sequence, sequence fl site ori of replication initiation, as well as Bg1II/Hindlll gene fragment, coding DNA of synthetic gene TβRII. Described is strain of bacteria Escherichia coli BL21 (DE3)/pET-32a/TβRII - producent of fusion protein thioredoxin- TβRII. Claimed is method of isolating target protein of human TβRII. Soluble fraction of cytoplasmatic proteins is isolated from strain-producent Escherichia coli BL21(DE3)/pET-32a/TβRII. Chromatographic purification with further renaturation of fusion protein thioredoxin- TβRII is carried out. After that, protein is incubated for 72 hours at 4°C in buffer, containing 0.5 M of arginine, in presence of oxidised and reduced glutathione in ratio 1:50 mM respectively. After that dialysis, decomposition of catalytic sununit with recombinant enteropeptidase and purification of target protein TβRII from thioredoxin are carried out.
EFFECT: invention makes it possible to obtain increased output of highly purified human TβRII protein with native N-end.
3 cl, 3 dwg, 1 tbl, 3 ex
| Title | Year | Author | Number |
|---|---|---|---|
| EXTRACTION METHOD OF RECOMBINANT PROTEINS | 2011 |
|
RU2499052C2 |
| METHOD OF PRODUCING HUMAN RECOMBINANT ANTI-TUMOUR PROTEIN DR5-B | 2018 |
|
RU2687435C1 |
| METHOD FOR MAKING MUTANT HUMAN TRAIL PROTEIN | 2009 |
|
RU2405038C1 |
| PLASMID VECTOR pE-Trx-Aur, STRAIN ESCHERICHIA COLI FOR EXPRESSION OF ANTIMICROBIAL PEPTIDE AURELIN AND METHOD OF SAID PEPTIDE OBTAINING | 2009 |
|
RU2412999C1 |
| RECOMBINANT PLASMID DNA OF PTRX-TEVRS-PTH CODING A HYBRID PROTEIN CAPABLE OF PROTEOLYTIC SPLITTING TO FORM A FRAGMENT OF ENDOGENOUS HUMAN PARATHYROID HORMONE (1-34), AN ESCHERICHIA COLI BL21(DE3)/PTRX-TEVRS-PTH STRAIN - PRODUCER OF SAID PROTEIN AND A METHOD OF PRODUCING RECOMBINANT PTH (1-34) | 2019 |
|
RU2700452C1 |
| RECOMBINANT PLASMID DNA pЕ-Trx-Lc-def, Escherichia coli STRAIN FOR EXPRESSION OF ANTIMICROBIAL PEPTIDE OF LENTIL DEFENSIN Lens culinaris AND METHOD FOR PRODUCING SAID PEPTIDE | 2010 |
|
RU2456345C1 |
| RECOMBINANT PLASMID DNA CODING FUSION PROTEIN L-HEP, ESCHERICHIA COLI STRAIN PRODUCER OF SAID PROTEIN AND METHOD OF PRODUCING RECOMBINANT PROTEIN | 2019 |
|
RU2716975C1 |
| METHOD FOR PURIFICATION OF RECOMBINANT PROTEIN CONTAINING MYELOPEPTIDES SEQUENCES | 2016 |
|
RU2630302C1 |
| RECOMBINANT PLASMID DNA pET22b(+)/slurp-2 CODING SLURP-2 PROTEIN, AND STRAIN OF BACTERIA Escherichia coli BL21(DE3) pET22b(+)/slurp-2 PRODUCER OF HUMAN SLURP-2 PROTEIN | 2015 |
|
RU2583307C2 |
| PLASMID VECTOR pE-Lc-LTP, Escherichia coli BACTERIUM STRAIN FOR EXPRESSION OF LIPID-TRANSPORTING LENTIL PROTEINS Lens culinaris AND METHOD FOR PRODUCING SAID PROTEINS | 2009 |
|
RU2415940C1 |
