title: Using synthetic biological parts and microbioreactors to explore the protein expression characteristics of Escherichia coli.
abstract: Synthetic biology has developed numerous parts for the precise control of protein expression. However, relatively little is known about the burden these place on a host, or their reliability under varying environmental conditions. To address this, we made use of synthetic transcriptional and translational elements to create a combinatorial library of constructs that modulated expression strength of a green fluorescent protein. Combining this library with a microbioreactor platform, we were able to perform a detailed large-scale assessment of transient expression and growth characteristics of two Escherichia coli strains across several temperatures. This revealed significant differences in the robustness of both strains to differing types of protein expression, and a complex response of transcriptional and translational elements to differing temperatures. This study supports the development of reliable synthetic biological systems capable of working across different hosts and environmental contexts. Plasmids developed during this work have been made publicly available to act as a reference set for future research.
authors.affiliation: DSM Biotechnology Center, P.O. Box 1, 2600 MA Delft, The Netherlands.
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Gorochowski
authors.forename: Eric
authors.initials: E
authors.lastname: van den Berg
authors.forename: Richard
authors.initials: R
authors.lastname: Kerkman
authors.forename: Johannes A
authors.initials: JA
authors.lastname: Roubos
authors.forename: Roel A L
authors.initials: RA
authors.lastname: Bovenberg
mesh_terms: D019149:Bioreactors; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D008954:Models, Biological; D011994:Recombinant Proteins / Q000378:metabolism*; D058615:Synthetic Biology; D049490:Systems Biology
chemical_list: D011994:Recombinant Proteins