| Popis: |
A chromatographic procedure was developed for the purification of ribosomes. Young pea plants were ground in a solvent containing polyethylene glycol, magnesium acetate, sucrose, and tris(hydroxymethyl)aminomethane acetic acid buffer (pH 7). The slurry was poured onto a column of cellulose and sand and the impurities washed out using a solvent containing magnesium acetate, sucrose, and tris(hydroxymethyl)-aminomethane—acetic acid buffer (pH7). The ribosomes were eluted by omitting magnesium acetate from the developing solvent. The effluent was centrifuged at 40 000 × g for 10 min. To achieve a higher degree of purification and concentration of the ribosomes, the chromatographic procedure was repeated with the supernatant. The solvent used for the grinding medium was used to eliminate more undesirable components from the eluate containing the ribosomes. This method completely separates chloroplasts and most of the other unwanted impurities from the ribosomes. The final preparation contained highly purified ribosomes, as evidenced by ultraviolet spectra, sedimentation constants and electron micrographs. |
