RECOMBINANT pET32A-TNF-Thy PLASMIDE PROVIDING THE SYNTHESIS OF THE FUSION PROTEIN Α-TUMOR NECROSIS FACTOR - THYMOSIN ALPHA 1, BACTERIAL STRAIN ESCHERICHIA COLI BL21(DE3)/pET32a-THF-Thy - PRODUCER OF THE HYBRID PROTEIN TNF-THY AND TNF-THY Thy Russian patent published in 2023 - IPC C12N15/00 C12N15/70 C12N15/12 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology, in particular to genetic engineering, and relates to a new bacterial strain of Escherichia coli BL21/pET32a-TNF-Thy, which can be used to obtain a hybrid protein alpha tumor necrosis factor - thymosin alpha 1 (TNF-Thy), used in the medical industry. The recombinant plasmid pET32a-TNF-Thy - 5929 base pairs (bp) in size, provides the synthesis of a protein having the amino acid sequence of SEQ ID NO: 1 with a size of 20.5 kDa, and contains structural elements: the T7 promoter and the 1acO operator, a synthetic nucleotide SEQ ID NO: 2 encoding TNF-Thy fusion protein, ampicillin resistance gene (AmpR) and bacterial ampicillin resistance gene promoter (AmpR promoter) for recombinant cell selection, origin of replication of bacteriophage f1 (f1 ori), gene for lactose the LacI repressor; and the bacterial promoter of the lactose repressor gene (Laclpromoter).

EFFECT: invention makes it possible to increase the efficiency of obtaining the TNF-Thy protein, namely, to increase the yield of the TNF-Thy hybrid protein from the strain biomass with a purity of at least 97% by creating a recombinant Escherichia coli strain - a producer of the TNF-Thy hybrid protein, obtained on the basis of the constructed recombinant plasmid, which provides an increased yield of the TNF-Thy hybrid protein by increasing the amount of biomass obtained during cultivation by a factor of 10 while maintaining the total protein content and the proportion of the TNF-Thy hybrid protein in the total cell protein.

8 cl, 3 dwg, 6 ex