RECOMBINANT PLASMID PET32-TREX VIC, WHICH PROVIDES FOR SYNTHESIS OF PROCHYMOSIN VICUGNA PACOS CHIMERIC PROTEIN, AND ESCHERICHIA COLI BL21(DE3)PLYSE PET32-TRX VIC-PRODUCER OF PROCHYMOSIN VICUGNA PACOS CHIMERIC PROTEIN Russian patent published in 2020 - IPC C12N15/64 C12N15/70 C12N9/64 C12N1/21 C12R1/19 

FIELD: biotechnology.

SUBSTANCE: group of inventions relates to a recombinant plasmid pET32-TrexVic, which provides synthesis of chimeric protein prochymosin Vicugna pacos, as well as a recombinant strain Escherichia coli, which produces said protein. Recombinant plasmid pET32-TrexVic has size of 6826 base pairs (bp) and provides synthesis of chimeric protein of prochymosin Vicugna pacos with size of 52 kDa in cells Escherichia coli BL21 (DE3) pLysE. Plasmid contains the structural elements essential for its functioning and expression of the target protein: a bacteriophage T7 promoter, a recombinant proichymosin Vicugna pacos gene encoding a target chimeric protein, a T7 RNA polymerase terminator, a thioredoxin sequence added to the nucleotide sequence of the prochymosin Vicugna pacos gene on the N-terminus, a bla (AmpR) sequence, protein coding beta-lactamase, sequence ColE1 (ori), providing replication of plasmid in E.coli strains. Recombinant protein gene of prochymosin Vicugna pacos has a nucleotide sequence SEQ ID NO: 1 nucleotide in position 5209 bp to nucleotide position 6649 bp, encoding the target chimeric protein of prochymosin Vicugna pacos with the full amino acid sequence SEQ ID NO: 2, built on the FauND I and HindIII restriction sites into the polyketer region of the pET32a vector containing the T7 phage TNA RNA polymerase promoter and terminator. What is also presented is Escherichia coli BL21 (DE3) pLysE/pET32-TrxVic strain producing prochymosin Vicugna pacos chimeric protein and obtained by transformation with recombinant plasmid pET32-TrexVic.

EFFECT: group of inventions provides high rate of refolding of target enzyme.

2 cl, 5 dwg, 2 tbl, 5 ex