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METHOD FOR PRODUCING Escherichia coli CELL FRACTION SHOWING PROTEASE INHIBITING ACTIVITY

机译:产生大肠杆菌细胞级分显示蛋白酶抑制活性的方法

摘要

FIELD: medicine.;SUBSTANCE: method for producing an Escherichia coli cell fraction showing protease inhibiting activity provides bacteria cells preservation in the presence of buffered 80-90% glycerine, processing by 3% triton X-100 for removing a cell membrane. Produced cytoplasm proteins are extracted by the increasing salt concentration, namely 0.14 M, 0.35 M; 2 M NaCl, 6 M guanidine hydrochloride with 0.1% β-mercaptoethanole. It is followed by affine sepharose 4B chromatography with immobilised trypsin and estimation of protease inhibiting activity in eluates.;EFFECT: invention can be used in analysis of molecular-genetic mechanisms of procaryote cell structure formation and roles of protein components in their organisation, and also genome remodelling nature that is necessary for disclosing of regulation pathways of mechanisms of macro-and microorganism action, and also search of new drug targets and development of ecologically safe new drugs.;3 tbl, 4 dwg
机译:领域:药物:产生具有蛋白酶抑制活性的大肠杆菌细胞级分的方法提供了在缓冲的80-90%甘油存在下细菌细胞的保存,通过3%triton X-100处理以去除细胞膜。通过增加盐浓度,即0.14 M,0.35 M,提取产生的细胞质蛋白。 2 M NaCl,6 M盐酸胍和0.1%β-巯基乙醇。随后进行仿射琼脂糖4B层析和固定化胰蛋白酶,并评估洗脱液中的蛋白酶抑制活性。效果:本发明可用于分析原核生物细胞结构形成的分子遗传机制以及蛋白质组分在其组织中的作用,以及基因组重塑性质,对于揭示宏观和微生物作用机制的调控途径,以及寻找新药靶标和开发生态安全的新药是必不可少的。3 tbl,4 dwg

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