Design of typical genes for heterologous gene expression

A publication (2022 | journal article) of the University of Göttingen

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​Design of typical genes for heterologous gene expression​
Simm, D. ; Popova, B. ; Braus, G. H. ; Waack, S.   & Kollmar, M. ​ (2022) 
Scientific Reports12(1).​

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Simm, Dominic 
Popova, Blagovesta 
Braus, Gerhard H. 
Waack, Stephan 
Kollmar, Martin 
Abstract Heterologous protein expression is an important method for analysing cellular functions of proteins, in genetic circuit engineering and in overexpressing proteins for biopharmaceutical applications and structural biology research. The degeneracy of the genetic code, which enables a single protein to be encoded by a multitude of synonymous gene sequences, plays an important role in regulating protein expression, but substantial uncertainty exists concerning the details of this phenomenon. Here we analyse the influence of a profiled codon usage adaptation approach on protein expression levels in the eukaryotic model organism Saccharomyces cerevisiae . We selected green fluorescent protein (GFP) and human α-synuclein (αSyn) as representatives for stable and intrinsically disordered proteins and representing a benchmark and a challenging test case. A new approach was implemented to design typical genes resembling the codon usage of any subset of endogenous genes. Using this approach, synthetic genes for GFP and αSyn were generated, heterologously expressed and evaluated in yeast. We demonstrate that GFP is expressed at high levels, and that the toxic αSyn can be adapted to endogenous, low-level expression. The new software is publicly available as a web-application for performing host-specific protein adaptations to a set of the most commonly used model organisms ( ).
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Scientific Reports 
Deutsche Forschungsgemeinschaft
Georg-August-Universität Göttingen
Open-Access-Publikationsfonds 2022



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