PET21A-PROCHYM RECOMBINATE PLASMID, PROVIDING SYNTHESIS OF CHEMERIC B BOS TAURUS PROXIMOSIN PROTEIN, AS WELL AS ESCHERICHIA COLI BL21 (DE3) PLYSE PET21A-PROCHYM - PRODUCER OF CHEMERIC B BOS TAURUS PROXIMOSIN PROTEIN

摘要

FIELD: chemistry.;SUBSTANCE: group of inventions relates to a recombinant plasmid providing the synthesis of a chimeric protein of B Bos Taurus prochymosin, and a recombinant strain of E. coli, a producer of chimeric protein of B Bos Taurus prochymosin in E. coli cells. Recombinant pET21a-ProChym plasmid of 6504 base pairs (bp) contains the following structural elements essential to its functioning and expression of the target protein: a bacteriophage T7 promoter that provides efficient transcription of the controlled mRNA; a recombinant gene of B Bos Taurus prochymosin encoding the target chimeric protein; T7 terminator, which is the untranslated termination region of the transcription of the bacterial operon providing an effective termination of transcription; the sequence of the T7 expression tag, added to the composition of the nucleotide sequence of the gene of B Bos Taurus prochymosin to the N-terminus, to increase the yield of the target protein; a bla sequence encoding a beta-lactamase protein, which is a selective marker for the transformation of E. coli strains; a ColE1 sequence that replicates the plasmid in E. coli strains. Recombinant gene of B Bos Taurus prochymosin has the nucleotide sequence of SEQ ID NO: 1 from the nucleotide at position 5205 bp. to the nucleotide at position 6347 bp, encoding the target chimeric protein of B Bos Taurus prochymosin with the complete amino acid sequence of SEQ ID NO: 2, embedded at the FauND I and Sfr274 I restriction sites into the region of the pET21a polylinker containing the T7 phage RNA polymerase promoter and terminator. Proposed recombinant strain of Escherichia coli BL21 (DE3) pLysE pET21a-ProChym is obtained by transformation with the recombinant plasmid pET21a-ProChym and deposited in the Collection of bacteria, bacteriophages and fungi of the State Research Center of Virology and Biotechnology VECTOR of the Federal Service for Supervision of Consumer Rights Protection and Human Well-Being under registration number B-1362.;EFFECT: group of inventions provides high production of pro-chymosin.;2 cl, 5 dwg, 6 ex, 2 tbl