【摘要】 本实验用改良密度梯度离心法对细胞各组份进行了逐级分离,成功地分离了微粒体并制备了较纯的粗面内质网和滑面内质网。在分级分离的各个阶段均进行了形态学观察及生化学检定,特别是最后得到了电镜证实。经分离纯化的滑面内质网杂蛋白显著减少而标志酶活性明显增高。在离心系统中,用KBr代替昂贵的CsCl,SDS代替去氧胆酸钠取得满意的分离效果。从而为滑面内质网及粗面内质网的分离和纯化建立了一种简单、经济、准确、可靠的方法。该技术的建立将为细胞的超微结构,生物工程,生物膜功能及酶促合成反应动力学等重要课题的深入研究提供了有效的手段。更多还原

【Abstract】 Cell fractions were isolated with method of density gradient centrifugation. Microsomes were separated and the purified smooth and rough endoplasmic reticula were obtained successfully. During each step, microscopic and biochemical examinations were performed for each fraction. Impure proteins in the endoplasmic reticulum decreased markedly after isolation and purification. Satisfactory results were obtained with KBr and SDS instead of CsCl and sodium deoxycholate in the centrifugation system. Thus, a simple, economical, accurate and reliable method to separate and purify microsomes was established. This technique makes it possible to study the cellular ultramicrostuctrure, biological engineering, functions of biological membrane and kinetics of enzymatic catalysis in synthetic reactions.更多还原