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Plasmid DNA Preparation> | … Plasmids and mRNA constructs DNA plasmids encoding nanoluciferase constructs were designed as follows. Nanoluciferase sequence from pNL1.1[Nluc] Vector (Promega) was inserted into pUC57(Kan) plasmid (Genscript), flanked by β-globin 5'- and 3'-UTRs … | Get A Quote |
Termination of protein biosynthesis is an essential step of gene expression, during which a complete functional protein is released from the ribosome. Premature or inefficient termination results in truncated, non-functional or toxic proteins that may cause disease. Indeed, more than 10% of human genetic diseases are caused by nonsense mutations leading to premature termination. Efficient and sensitive approaches are required to study eukaryotic termination mechanisms and to identify potential therapeutics that modulate termination. Canonical radioactivity-based termination assays are complex, report on a short peptide release, and are incompatible with high-throughput screening. Here we describe a robust and s... More