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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。
In this experiment, the cells were separated step by step by modified density gradient centrifugation, the microsomes were successfully separated and the pure rough endoplasmic reticulum and smooth endoplasmic reticulum were prepared. Morphological and biochemical tests were performed at all stages of fractionation, especially with electron microscopy. The isolated and purified slide endoplasmic reticulum heterozygote significantly reduced the marked enzyme activity was significantly increased. In centrifuge systems, KBr was used instead of expensive CsCl, and SDS was used in place of sodium deoxycholate to achieve satisfactory separation. Thus, a simple, economical, accurate and reliable method for the separation and purification of endoplasmic reticulum and rough endoplasmic reticulum was established. The establishment of this technology will provide an effective means for further research on important topics such as cell ultrastructure, bioengineering, biofilm function and enzymatic synthesis reaction kinetics.