Fabrication and Cross-Chassis-Characterisation of Artificial Promoters and 5´ Untranslated Regions in Vibrio Natriegens and Escherichia coli

Vibrio natriegens is a rising star on the horizon of microbial gene expression chassis, growing even faster than model organism Escherichia coli. To express genes in Vibrio natriegens, we currently use genetic elements optimized for Escherichia coli because few of Vibrio natriegens’ native genetic elements are known and characterized. This means that Vibrio natriegens’ full potential as a gene expression chassis remains unknown. Characterizing native genetic elements is a cost- and time-consuming process. To reduce time and cost, we used the Gene Expression Engineering (GeneEE) method to efficiently create and characterize artificial genetic elements (promoters and 5´ Untranslated Regions) that lead to expression of Green Fluorescent Protein and mCherry in a wide range of expression levels. We characterized the artificial genetic elements in Vibrio natriegens and Escherichia coli and found that some of the artificial genetic elements lead to high expression in Vibrio natriegens but not in Escherichia coli. With our study we expand the pool of known and characterized genetic elements in Vibrio natriegens, which can be used for many different biotechnological applications utilizing Vibrio natriegens as a gene expression chassis.