Polysome Profiling and Metabolic Labeling Methods to Measure Translation in Trypanosoma brucei
Polysome Profiling and Metabolic Labeling Methods to Measure Translation in Trypanosoma brucei
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DOI：
10.1007/978-1-0716-0294-2_7
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发表时间：
2020-01-01
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影响因子：
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通讯作者：
Clayton, Christine
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文献类型：
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作者：
Bajak, Kathrin;Clayton, Christine
细胞中产生的蛋白质的量不仅取决于相应的 mRNA 水平，还取决于 mRNA 的翻译效率。非常强大的全转录组方法可用于分析每个 mRNA 上核糖体的密度和多肽延伸的速率。然而，对于许多研究问题，更简单、更便宜的方法更合适。在这里，我们描述了两种评估细胞一般翻译状态的方法：通过蔗糖密度梯度离心进行多核糖体分析和使用放射性氨基酸进行代谢标记。这两种方法也可用于检查单个 mRNA 的翻译。
The amount of a protein that is made in a cell is determined not only by the corresponding mRNA level but also by the efficiency with which the mRNA is translated. Very powerful transcriptome-wide methods are available to analyze both the density of ribosomes on each mRNA and the rate at which polypeptides are elongated. However, for many research questions, simpler, less expensive methods are more suitable. Here we describe two methods to assess the general translation status of cells: polysome profiling by sucrose density gradient centrifugation and metabolic labeling using radioactive amino acids. Both methods can also be used to examine translation of individual mRNAs.