Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, Canada; Department of Mathematics, University of British Columbia, Vancouver, BC, Canada.
School of Computer Science, University of Waterloo, Waterloo, ON, Canada; Department of Computer Science, University of British Columbia, Vancouver, BC, Canada.
STAR Protoc. 2022 Aug 18;3(3):101605. doi: 10.1016/j.xpro.2022.101605. eCollection 2022 Sep 16.
Ribosome profiling is a powerful technique which maps the distribution of ribosomes along mRNAs to analyze translation genome-wide. Ribosome density can be affected by multiple factors, such as changes to translation initiation or elongation rates. We describe the application of a metric for identifying genes rate-limited by these rates by analyzing the relative distribution of ribosome footprints along transcripts. This protocol also details two sample analyses comparing gene translation efficiencies and the distribution of ribosome densities on downloadable datasets. For complete details on the use and execution of this protocol, please refer to Flanagan et al. (2022).
核糖体图谱分析是一种强大的技术,它可以绘制核糖体在 mRNA 上的分布情况,从而对整个基因组的翻译进行分析。核糖体密度可能会受到多种因素的影响,例如翻译起始或延伸速度的变化。我们通过分析核糖体足迹在转录本上的相对分布,描述了一种通过分析相对分布来识别受这些速率限制的基因的度量方法。该方案还详细介绍了两种比较基因翻译效率和核糖体密度分布的样本分析,这些分析可在可下载的数据集上进行。有关该方案使用和执行的完整详细信息,请参阅 Flanagan 等人(2022 年)。