METHOD OF PRODUCTION OF NUCLEAR PROTEINASES OF NON-HISTONE AND HISTONE PLANT PROTEINS

摘要

FIELD: biotechnology.;SUBSTANCE: method of production of nuclear proteinases of non-histone and histone plant proteins, in which initially in certain time intervals from the start of soaking 0 h, 3 h, 6 h, 9 h, 12 h, 15 h, 18 h, 21 h, separation of corcules from endosperm is carried out the followed by preservation of the corcules in the buffered 80-90% glycerol at -25°C, cell nuclei are isolated, the nuclear fractions are extracted with increasing concentrations of 0.14 M, 0.35 M, 2 M sodium chloride and 6 M guanidine hydrochloride with 0.1% β-mercaptoethanol, the histone and non-histone proteins are isolated from the above fractions using ion exchange chromatography with amberlite IRC-50 in the discontinuous gradient of guanidine hydrochloride: 6%, 8.9%, 10.6%, 13%, 40% on 0.1 M potassium-phosphate buffer pH 6.8, the resulting proteins are passed using the method of affinity chromatography through the column with sepharose 4B with immobilised trypsin inhibitor followed by evaluation of the proteolytic activity.;EFFECT: invention can be used to analyse the molecular-genetic mechanisms of formation of structures of the cell nucleus, which is necessary to obtain additional information in the development and construction of computer models of organisation of gene and epigene control networks.;4 dwg