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Author:
Date:
2014-04-05
[Abstract] UPF1, an RNA helicase and a core factor of nonsense-mediated mRNA decay (NMD), interacts with RNA independently of the sequence context. To investigate the influence of translation on the association of UPF1 with specific reporter transcripts, UPF1 RNA immunoprecipitations (RIPs) are performed from Hela cells that either express a normally translated immunoglobulin-µ (Ig-µ) reporter (mini µ) or a version with a stable stem loop in the 5' UTR (SL mini µ) that efficiently inhibit translation initiation (Zund et al., 2013). Both the cloning of the SL mini µ reporter construct and the ...
[摘要] UPF1，RNA解旋酶和无义介导的mRNA衰变（NMD）的核心因素，独立于序列上下文与RNA相互作用。 为了研究翻译对UPF1与特异性报告基因转录物的关联的影响，从表达正常翻译的免疫球蛋白-μ（Ig-μ）报道分子（微型μ）的Hela细胞或其中的一个版本进行UPF1RNA免疫沉淀（RIP） 在5'UTR（SL miniμ）中的稳定茎环，其有效抑制翻译起始（Zund等人，2013）。 详细描述了SL miniμ报告构建体的克隆和UPF1 RIP实验
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