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Non-programmed transcriptional frameshifting is common and highly RNA polymerase type-dependent

机译:非程序性转录移码是常见的且高度依赖RNA聚合酶类型

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摘要

BackgroundThe viral or host systems for a gene expression assume repeatability of the process and high quality of the protein product. Since level and fidelity of transcription primarily determines the overall efficiency, all factors contributing to their decrease should be identified and optimized. Among many observed processes, non-programmed insertion/deletion (indel) of nucleotide during transcription (slippage) occurring at homopolymeric A/T sequences within a gene can considerably impact its expression. To date, no comparative study of the most utilized Escherichia coli and T7 bacteriophage RNA polymerases (RNAP) propensity for this type of erroneous mRNA synthesis has been reported. To address this issue we evaluated the influence of shift-prone A/T sequences by assessing indel-dependent phenotypic changes. RNAP-specific expression profile was examined using two of the most potent promoters, ParaBAD of E. coli and φ10 of phage T7.
机译:背景技术用于基因表达的病毒或宿主系统假定过程的可重复性和蛋白质产物的高质量。由于转录的水平和保真度主要决定了整体效率,因此应确定和优化所有有助于降低转录效率的因素。在许多观察到的过程中,在基因内的同聚A / T序列处发生的转录(滑移)过程中核苷酸的非编程插入/缺失(插入/缺失)会极大地影响其表达。迄今为止,尚未报道对这种错误的mRNA合成最常用的大肠杆菌和T7噬菌体RNA聚合酶(RNAP)倾向的比较研究。为了解决这个问题,我们通过评估依赖于indel的表型变化来评估易移位A / T序列的影响。使用两个最有效的启动子,即大肠杆菌的ParaBAD和噬菌体T7的φ10,检查了RNAP特异性表达谱。

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