دورية أكاديمية
Performing Ribosome Profiling to Assess Translation in Vegetative and Meiotic Yeast Cells.
| العنوان: | Performing Ribosome Profiling to Assess Translation in Vegetative and Meiotic Yeast Cells. |
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| المؤلفون: | Powers EN; Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA, USA. epow@berkeley.edu., Brar GA; Department of Molecular and Cell Biology, University of California at Berkeley, Berkeley, CA, USA. gabrar@berkeley.edu. |
| المصدر: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2252, pp. 89-125. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Totowa, NJ : Humana Press Original Publication: Clifton, N.J. : Humana Press, |
| مواضيع طبية MeSH: | RNA, Messenger/*genetics , Ribosomes/*metabolism , Saccharomycetales/*physiology , Sequence Analysis, RNA/*methods, Centrifugation, Density Gradient ; Freezing ; Fungal Proteins/metabolism ; Gene Expression Profiling ; High-Throughput Nucleotide Sequencing ; Meiosis ; Protein Biosynthesis ; Saccharomycetales/genetics |
| مستخلص: | Ribosome profiling, first developed in 2009, is the gold standard for quantifying and qualifying changes to translation genome-wide (Ingolia et al., Science, 2009). Though first designed and optimized in vegetative budding yeast, it has since been modified and specialized for use in diverse cellular states in yeast, as well as in bacteria, plants, human cells, and many other organisms (Ingolia et al. Science, 2009, reviewed in (Ingolia et al., Cold Spring Harb Perspect Biol, 2019; Brar and Weissman, Nat Rev Mol Cell Biol, 2015)). Here we report the current ribosome profiling protocol used in our lab to study genome-wide changes to translation in budding yeast undergoing the developmental process of meiosis (Brar et al., Science, 2012; Cheng et al., Cell, 2018). We describe this protocol in detail, including the following steps: collection and flash freezing samples, cell lysis and extract preparation, sucrose gradient centrifugation and monosome collection, RNA extraction, library preparation, and library quality control. Almost every step presented here should be directly applicable to performing ribosome profiling in other eukaryotic cell types or cell states. |
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| معلومات مُعتمدة: | R35 GM134886 United States GM NIGMS NIH HHS |
| فهرسة مساهمة: | Keywords: Cellular development; Gene expression; Meiosis; Ribosome profiling; Translation; Yeast |
| المشرفين على المادة: | 0 (Fungal Proteins) 0 (RNA, Messenger) |
| تواريخ الأحداث: | Date Created: 20210325 Date Completed: 20210615 Latest Revision: 20211015 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC8513646 |
| DOI: | 10.1007/978-1-0716-1150-0_4 |
| PMID: | 33765272 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1940-6029 |
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| DOI: | 10.1007/978-1-0716-1150-0_4 |