免疫沉淀
核糖核蛋白
翻译(生物学)
异质核核糖核蛋白
核糖核酸
真核翻译
细胞生物学
计算生物学
生物
蛋白质生物合成
信使核糖核酸
化学
平动调节
EIF4E公司
RNA结合蛋白
分子生物学
基因
遗传学
作者
Chris Lucchesi,Shakur Mohibi,Xinbin Chen
出处
期刊:Methods in molecular biology
日期:2021-01-01
卷期号:: 73-79
被引量:1
标识
DOI:10.1007/978-1-0716-1217-0_5
摘要
Eukaryotic mRNAs are bound by a multitude of RNA binding proteins (RBPs) that control their localization, transport, and translation. Measuring the rate of translation of mRNAs is critical for understanding the factors and pathways involved in gene expression. In this chapter, we present a method to compare the rate of translation of individual mRNA species based on the fraction of mRNA bound by a specific ribonucleoprotein involved in the general translation machinery. The ribonucleoprotein complex is immunoprecipitated using an antibody for the RBP, followed by RT-PCR to semi-quantitatively determine the amount of an individual mRNA fraction bound by a translation regulating protein such as eIF4E.
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