FIELD: genetic engineering.
SUBSTANCE: invention relates to the field of genetic engineering and molecular biology, in particular to the recombinant plasmid pET32v11-Cre, which provides synthesis of the recombinant protein 6His-S-NLS-Cre. The invention also discloses a strain of Escherichia coli BL21(DE3)/pET32v11-Cre cells and a method for obtaining the specified 6His-S-NLS-Cre protein using the specified strain. The plasmid according to the invention provides synthesis of the specified recombinant protein in Escherichia coli cells due to the location of the Cre recombinase gene in the 3’ region relative to the T7 promoter and lac operator and in the 5’ region relative to the T7 terminator and was obtained by cloning the cDNA of Cre recombinase into the pET32v11 vector, so that the cDNA of Cre recombinase with N-terminal NLS was in the same reading frame with 6-histidine and S-epitopes and amino acid sequences of thrombin, enterokinase, and TEV protease cleavage sites. It can be used for the production of recombinant site-specific recombinase Cre.
EFFECT: present invention makes it possible to obtain a catalytically active Cre recombinase with a high yield.
4 cl, 6 dwg, 5 ex
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Authors
Dates
2021-12-13—Published
2020-12-30—Filed