FIELD: medicine.
SUBSTANCE: what is presented is a method of preparative recovery of basic proteins from supramolecular structures of Escherichia coli growing population. The Escherichia coli cells are preserved in buffered 80-90% glycerol at -25°C. Then the sediment cells are washed in 3% triton X-100. It is followed by sediment extraction in salts of increasing concentrations: 0.14 M, 0.35 M; 2 M NaCl, 6 M in guanidine hydrochloride with 0.1% β-mercaptoethanol. Basic proteins are recovered from the prepared fractions by means of ion-exchange chromatography with amberlite resin IRC-50 of discontinuous gradient of guanidine hydrochloride: 6%, 8.9%, 10.6%, 13% on 0.1 M potassium phosphate buffer pH 6.8.
EFFECT: method enables producing fractions enriched by basic proteins with the use of a microamount of protein of Escherichia coli cell suprastructures.
7 dwg, 1 ex
