在酿酒酵母中通过 TIS 分析进行翻译起始位点的全局作图。
Global mapping of translation initiation sites by TIS profiling in budding yeast.
机构信息
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
California Institute for Quantitative Biosciences (QB3), University of California, San Francisco, CA 94158, USA.
出版信息
STAR Protoc. 2020 Dec 30;2(1):100250. doi: 10.1016/j.xpro.2020.100250. eCollection 2021 Mar 19.
Translation initiation site (TIS) profiling allows for the genome-wide identification of TISs by exclusively capturing mRNA fragments within ribosomes that have just completed translation initiation. It leverages translation inhibitors, such as harringtonine and lactimidomycin (LTM), that preferentially capture ribosomes at start codon positions, protecting TIS-derived mRNA fragments from nuclease digestion. Here, we describe a step-by-step protocol for TIS profiling in LTM-treated budding yeast that we developed to identify TISs and open reading frames in vegetative and meiotic cells. For complete details on the use and execution of this protocol, please refer to Eisenberg et al. (2020).
翻译起始位点（TIS）谱分析通过专门捕获刚刚完成翻译起始的核糖体中的 mRNA 片段，实现了全基因组范围内 TIS 的识别。它利用了翻译抑制剂，如哈林通宁和乳酰亚胺霉素（LTM），这些抑制剂优先在起始密码子位置捕获核糖体，从而保护 TIS 衍生的 mRNA 片段免受核酸酶的消化。在这里，我们描述了在 LTM 处理的出芽酵母中进行 TIS 谱分析的逐步方案，我们开发该方案是为了鉴定营养细胞和减数分裂细胞中的 TIS 和开放阅读框。有关此方案的使用和执行的完整详细信息，请参阅 Eisenberg 等人。（2020）。
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