The regulation of gene expression occurs through complex relationships between transcription, processing, turnover, and translation, which are only beginning to be elucidated. We know that at least for certain messenger (m) RNAs, processing, modifications, and sequence elements can greatly influence their translational output through recognition by translation and turn-over machinery. Recently, we and others have combined high-throughput sequencing technologies with traditional biochemical methods of studying translation to extend our understanding of these relationships. Additionally, there is growing importance given to how these processes may be regulated across varied cell types as a means to achieve tissue-specific expression of proteins. Here, we provide an in-depth methodology for polysome profiling to dissect the composition of mRNAs and proteins that make up the translatome from both whole tissues and a specific cell type isolated from mammalian tissue. Also, we provide a detailed computational workflow for the analysis of the next-generation sequencing data generated from these experiments.

中文翻译：

---

来自哺乳动物组织的细胞类型特异性多核糖体分析

基因表达的调控通过转录、加工、转换和翻译之间的复杂关系发生，这些关系才刚刚开始阐明。我们知道，至少对于某些信使 (m) RNA，加工、修饰和序列元素可以通过翻译和翻转机制的识别极大地影响它们的翻译输出。最近，我们和其他人将高通量测序技术与研究翻译的传统生化方法相结合，以扩展我们对这些关系的理解。此外，作为实现蛋白质组织特异性表达的一种手段，如何在不同细胞类型中调节这些过程变得越来越重要。这里，我们为多核糖体分析提供了一种深入的方法，以剖析构成整个组织和从哺乳动物组织中分离的特定细胞类型的翻译组的 mRNA 和蛋白质的组成。此外，我们还提供了详细的计算工作流程，用于分析从这些实验中生成的下一代测序数据。