METHOD FOR ISOLATION OF PROTEIN 39 KDA FROM MELIODOSIS PATHOGEN EXTERNAL MEMBRANE
FIELD: medicine, preparative biochemistry, proteins, microbiology. SUBSTANCE: invention describes method for isolation of membrane protein 39 kDa from meliodosis pathogen (Burkholderia pseudomallei). Isolation of protein 39 kDa involves steps of microorganisms culturing, biomass washing off, treatme...
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Zusammenfassung: | FIELD: medicine, preparative biochemistry, proteins, microbiology. SUBSTANCE: invention describes method for isolation of membrane protein 39 kDa from meliodosis pathogen (Burkholderia pseudomallei). Isolation of protein 39 kDa involves steps of microorganisms culturing, biomass washing off, treatment of cellular suspension by ultrasonic oscillation in ice, centrifugation of suspension at 1 500g, two-fold centrifugation at 100 000g to isolate the total preparation of cellular membranes (EM) in step-by-step gradient of sucrose density (55%, 43%, 35%, 19%) at 100 000g to isolate external membranes, the following gel- -chromatography EM on TSK HW-60F (Toyopearl), preparative electrophoresis of the 1-st elution peak and electroelution of protein 39 kDa. Invention provides preparing "major" protein 39 kDa EM from B. pseudomallei representing highly purified protein with its complete retained antigenic activity and useful for investigation of its immunobiological properties, the development of effective specific identifying immunological test-systems and construction of immunoprophylactic preparations. EFFECT: improved isolating method, valuable properties of proteon. 3 dwg, 5 ex |
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