Ribosome purification from Escherichia coli by ultracentrifugation

Yi Cui , Xinjie Chen , Ze Wang , Yuan Lu
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Abstract

With more and more researchers conducting extensive research on all aspects of ribosomes, how to extract ribosomes with good effect and high activity has become a fundamental problem. In this article, Escherichia coli A19, MRE600, and JE28 cells often mentioned in the literature and ordinary E. coli BL21(DE3) cells were used to extract ribosomes by ultracentrifugation. The purpose was to study whether the ultracentrifugation method can be applied to extract effective ribosomes, and whether the ribosome extracts from different cells were different. The extracted ribosomes were validated by RNA electrophoresis, SDS-PAGE, PURE system, and mass spectrometry. The validation experiment results showed that ribosomes from these four cells had different effects. The success of the experiment confirmed that effective ribosomes could be extracted from E. coli by ultracentrifugation, which laid a good foundation for researchers to carry out further applications on ribosomes.

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超离心法纯化大肠杆菌核糖体
随着越来越多的研究者对核糖体的各个方面进行广泛的研究,如何提取出效果好、活性高的核糖体已成为一个根本性的问题。本文采用文献中经常提到的大肠埃希菌A19、MRE600、JE28细胞和普通的大肠埃希菌BL21(DE3)细胞进行超离心提取核糖体。目的是研究超离心方法是否可以提取有效的核糖体,不同细胞的核糖体提取物是否不同。提取的核糖体经RNA电泳、SDS-PAGE、PURE系统和质谱验证。验证实验结果表明,这四种细胞的核糖体具有不同的作用。实验的成功证实了利用超离心技术可以从大肠杆菌中提取有效的核糖体,为研究人员开展核糖体的进一步应用奠定了良好的基础。
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