Open AccessInfluence of the spacer region between the Shine–Dalgarno box and the start codon for fine‐tuning of the translation efficiency in Escherichia coli
Author(s) -
Komarova Ekaterina S.,
Chervontseva Zoya S.,
Osterman Ilya A.,
Evfratov Sergey A.,
Rubtsova Maria P.,
Zatsepin Timofei S.,
Semashko Tatiana A.,
Kostryukova Elena S.,
Bogdanov Alexey A.,
Gelfand Mikhail S.,
Dontsova Olga A.,
Sergiev Petr V.
Publication year - 2020
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/1751-7915.13561
Subject(s) - start codon , shine dalgarno sequence , eukaryotic translation , translational efficiency , translation (biology) , biology , protein biosynthesis , genetics , stop codon , eukaryotic translation initiation factor 4 gamma , gene , computational biology , messenger rna
Summary Translation efficiency contributes several orders of magnitude difference in the overall yield of exogenous gene expression in bacteria. In diverse bacteria, the translation initiation site, whose sequence is the primary determinant of the translation performance, is comprised of the start codon and the Shine–Dalgarno box located upstream. Here, we have examined how the sequence of a spacer between these main components of the translation initiation site contributes to the yield of synthesized protein. We have created a library of reporter constructs with the randomized spacer region, performed fluorescently activated cell sorting and applied next‐generation sequencing analysis (the FlowSeq protocol). As a result, we have identified sequence motifs for the spacer region between the Shine–Dalgarno box and AUG start codon that may modulate the translation efficiency in a 100‐fold range.