RECOMBINANT PLASMID DNA pPHOFus CODING ANTHRAX LETHAL FACTOR SUBSTRATUM FUSED WITH ALKALINE PHOSPHATASE Escherichia coli, AND Escherichia coli BL-PHOFus STRAIN PRODUCING ANTHRAX LETHAL FACTOR PROTEIN SUBSTRATUM AS INGREDIENT OF ALKALINE PHOSPHATASE Escherichia coli

摘要

FIELD: medicine.;SUBSTANCE: recombinant plasmid pETPHOFus enabling synthesis of an anthrax lethal factor substratum as a part of one polypeptide with a mutated reporter enzyme of alkaline phosphatase Escherichia coli. Said fused polypeptide represents a new high-sensitivity anthrax lethal factor substratum. Besides, an Escherichia coli BL-PHOFus strain being a producer of the fused polypeptide is offered. The strain provides a no-fermentation high-yield synthesized protein not less than 30 mg in 1 litre of a liquid culture. The prepared anthrax lethal factor substratum shows hypersensitiveness and high specificity to LF splitting and is able to detect proteolytic activity of the anthrax lethal factor in concentration up to 1 pM.;EFFECT: invention can find application in clinical recognition of anthrax disease; for detecting hot spots of anthrax, for screening the compound libraries and searching anthrax lethal factor activity inhibitors.;2 cl, 4 dwg, 4 ex