Team:UQ-Australia/Parts

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The human circadian rhythm drives many important processes in the body in accordance with the sleep/wake cycle. A characteristic of this biological clock is the periodic oscillation of gene expression. Current parts in the Registry designed to regulate periodic oscillations of gene expression have shown limited success.

Here we demonstrate a biological clock being standardised as a set of BioBrick parts.

Our network is controlled by an engineered promoter, Plac/ara, which features both an activator and a repressor domain. This controls the production of downstream genes to activate other inducible promoters, pBAD and GlnAp2, eventually leading to the production of a repressor protein, lacI, which inhibits Plac/ara, resulting in oscillatory expression. This project shows the feasibility of standardising the biological clock in E. coli and grounds further development for applications in regulated drug/hormone delivery and ion channel control.

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Parts

Promoter image.png Plac/ara

A multi-domain promoter that is self-inducible by arabinose. Gene expression regulated by LacI binding to special operator sites in the promoter

125x125px glnG

Encodes for DNA-binding protein(NR1), the activity of which depends on its phosphorylation. Drives expression of genes under GlnAp2 promoter

125x125px araC

Encodes for a positive regulatory protein for L-arabinose utilization in E.coli

Promoter image.png pBAD

L-arabinose inducible promoter

125x125px LacI

DNA-binding transcriptional inhibitor for lac-operon

Promoter image.png GlnAp2

Promoter driven by phosphorylated GlnG(NR1)