Influence of the spacer region between the Shine-Dalgarno box and the start codon for fine-tuning of the translation efficiency in Escherichia coliстатья

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[1] Influence of the spacer region between the shine-dalgarno box and the start codon for fine-tuning of the translation efficiency in escherichia coli / E. S. Komarova, Z. S. Chervontseva, I. A. Osterman et al. // Microbial Biotechnology. — 2020. Translation efficiency contributes several orders ofmagnitude difference in the overall yield of exoge-nous gene expression in bacteria. In diverse bacte-ria, the translation initiation site, whose sequence isthe primary determinant of the translation perfor-mance, is comprised of the start codon and the Shine–Dalgarno box located upstream. Here, wehave examined how the sequence of a spacerbetween these main components of the translationinitiation site contributes to the yield of synthesizedprotein. We have created a library of reporter con-structs with the randomized spacer region, per-formedfluorescently activated cell sorting andapplied next-generation sequencing analysis (theFlowSeq protocol). As a result, we have identifiedsequence motifs for the spacer region between theShine–Dalgarno box and AUG start codon that maymodulate the translation efficiency in a 100-fold range. [ DOI ]

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