METHOD OF PRODUCING RECOMBINANT BRUCELLOSIS PROTEIN rLptD Russian patent published in 2024 - IPC C12N15/63 C12N15/70 

FIELD: biotechnology.

SUBSTANCE: described is a method of producing recombinant brucellosis protein rLptD. Method includes constructing a gene of rLptD protein with replacement of nucleotides "gc" with "cg" in position 437 bp, creation of recombinant plasmid, transformation of cells of E. coli Shuffle T7 with expression plasmid DNA, culturing the obtained strain, obtaining a cytoplasmic fraction of producer bacteria by treating the cells with lysozyme, followed by centrifugation. This is followed by purification of the rLptD protein by metal-chelate chromatography to obtain a homogeneous final preparation of the rLptD protein with a yield of at least 0.2 g of protein per liter of the bacterial culture, having 98% purity according to electrophoresis and densitometry and containing bacterial endotoxin in an amount of not more than 5 IU/mg.

EFFECT: creation of a method for production and purification of homogeneous recombinant protein rLptD with purity of 98% from bacterial cytoplasm, content of bacterial endotoxin is not more than 5 IU/mg and output of 0,2 g per liter of bacterial culture, applicable to obtain large amounts of this protein, suitable for use in biotechnology and pharmacology.

1 cl, 4 dwg, 2 tbl, 6 ex