METHOD TO OBTAIN CHEMERIC RECOMBINANT PROTEIN fliC:pagN Russian patent published in 2017 - IPC C07K14/255 

FIELD: biotechnology.

SUBSTANCE: to implement the method, a recombinant plasmid pETfliCpagN is constructed, then Escherichia coli BL21 (DE3) cells with the said plasmid and the resulting BL-fliCpagN strain is cultured at a temperature of 37°C. Next, a periplasmic fraction of producer bacteria is prepared by cells treatment with lysozyme in a 20% sucrose solution, followed by centrifugation, the chemeric fliC:pagN by metal chelate, anion exchange, and gel filtration chromatography to obtain a homogeneous final preparation of fliC:pagN protein. The recombinant pETfliCpagN plasmid contains the T7 bacteriophage promoter and carries sequences encoding fliC proteins, including the leader sequence of this protein, and pagN S. typhimurium, combined with a short linker sequence GSVDSSSGGN, and fused at the 3'-end with a synthetic sequence encoding six histidins.

EFFECT: method allows to synthesise this protein in soluble form with a native N-terminal amino acid sequence, yielding at least 0,3 g of protein from a litre of bacterial culture with 98 percent purity.

7 dwg, 4 ex