METHOD FOR OBTAINING OF RECOMBINANT PEPTIDOGLYCAN-ASSOCIATED LIPOPROTEIN (PAL) LEGIONELLA PNEUMOPHILA Russian patent published in 2017 - IPC C12N15/09 C12N15/31 C12N15/64 C12N15/70 C07K14/195 

FIELD: biotechnology.

SUBSTANCE: method for obtaining of recombinant peptidoglycan-associated lipoprotein (PAL) Legionella pneumophila is proposed. A pET22b(+)-PAL plasmid with length of 6184 is constructed, bearing an expression construct for production of a chimeric polypeptide containing a sequence encoding a PAL protein fused at the 5'-end with a synthetic sequence encoding six histidins and a SUMO peptide. E. coli BL21 (DE3) cells are transformed with the expression plasmid DNA pET22b(+)-PAL. The resulting BL21(DE3)-PAL strain is cultured at 37°C with production of PAL protein for 3 hours. The HIS-SUMO-PAL protein is purified from the soluble fraction of bacterial lysate by metal-chelate chromatography. The HIS-SUMO peptide is cleaved by treatment with SUMO protease to restore the native structure of the PAL protein. Using ion-exchange and gel filtration chromatography, a homogeneous preparation of the PAL protein is obtained.

EFFECT: invention provides a PAL protein having 98 percent purity based on electrophoresis and densitometry data, yielding at least 300 mg per litre of bacterial culture without fermentation, which can be used in development of diagnostic preparations for legionellosis determination.

5 dwg, 3 ex