WAY OF RECEPTION FUNCTIONALLY ACTIVE RECOMBINANT PROTEIN OF ULCER (LF) LETHAL FACTOR, RECOMBINANT PLASMIDE DNA pETHIS-LF CODING ACTIVE PROTEIN LF; H STRAIN ESCHERICHIA COLI BL-HISLF PRODUCING ACTIVE PROTEIN OF MALIGNANT ANTHRAX LETHAL FACTOR

摘要

FIELD: medicine; microbiology. ^ SUBSTANCE: way is intended for reception of functionally active LF form, the basic toxic protein defining cellular disturbances, leading to death of an organism at infection with a malignant anthrax bacterium. For realisation of the way a recombinant plasmid pETHIS-LF (7816 items) is designed, containing a full-size gene of the lethal factor (LF) of malignant anthrax under the control of the promotor of bacteriophage T7 and to a determinant of ampicillin tolerance. The plasmide provides effective synthesis of LF protein of malignant anthrax merged with sequence of six Histidinums for clearing with the metal-chelate chromatography. The strain Escherichia coli BL-HISLF is designed using transformation of the specified plasmid DNA in the strain E.coli BL21 (DE3), synthesizing active LF protein. The target product is separated with the way including clearing on a metal-chelate sorbent with the subsequent additional clearing of the LF protein by gel-filtration. ^ EFFECT: reception of active recombinant protein LF on the simplified technology and with a high output of synthesised protein of the lethal factor. ^ 3 cl, 3 dwg, 3 ex