Unigems: plasmids and parts to facilitate teaching on assembly, gene expression control and logic in E. coli

Author:

Siddall Alex123,Williams Abbie Ann3,Sanders Jason4,Denton Jai A.51,Madden Dean6,Schollar John6,Bryk Jarosław36ORCID

Affiliation:

1. Genomics and Regulatory Systems Unit, Okinawa Institute of Science and Technology Graduate University, Okinawa, Japan

2. School of Biological Sciences, University of East Anglia, Norwich, UK

3. School of Applied Sciences, University of Huddersfield, Huddersfield, UK

4. School of Art, Design and Architecture, University of Huddersfield, Huddersfield, UK

5. Institute of Vector-borne Disease, Monash University, Clayton, Australia

6. National Centre for Biotechnology Education, University of Reading, Reading, UK

Abstract

Synthetic biology enables the creative combination of engineering and molecular biology for exploration of fundamental aspects of biological phenomena. However, there are limited resources available for such applications in the educational context, where straightforward setup, easily measurable phenotypes and extensibility are of particular importance. We developed unigems, a set of ten plasmids that enable classroom-based investigation of gene-expression control and biological logic gates to facilitate teaching synthetic biology and genetic engineering. It is built on a high-copy plasmid backbone and is easily extensible thanks to a common primer set that facilitates Gibson assembly of PCR-generated or synthesized DNA parts into the target vector. It includes two reporter genes with either two constitutive (high- or low-level) or two inducible (lactose- or arabinose-) promoters, as well as a single-plasmid implementation of an AND logic gate. The set can readily be employed in undergraduate teaching settings, during outreach events and for training of iGEM teams. All plasmids have been deposited in Addgene.

Funder

FP7 People: Marie-Curie Actions

Wellcome Trust

Publisher

Microbiology Society

Subject

Microbiology (medical),Microbiology

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