Team:NYMU-Taipei/Project/Time Regulation/Reloxilator

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Our Reloxilator (Relaxation Oscillator): A four-part system consisting of an oscillator, a synchronizer, a tuner, and a reporter.

NYMU reloxilator.png

Description

The oscillator is a two-component relaxation oscillator. The first component consists of the PRE promoter and the gene CII in a positive feedback loop, and both originating from from the λ phage. The second component uses the HtlB gene from E. coli attached to the same promoter as the first component. The HtlB gene produces HtlB proteins that degrade CII proteins.

The synchronizer allows near-full synchronization between different cells after about two time periods of oscillation1. The main parts of the synchronizer is made up of the PLux promoter, and the LuxI and LuxR genes, all coming from the Vibrio Fischeri organism. PRE and the CII gene are the same as the parts used in the oscillator, tying the oscillator and synchronizer together.

The Tuner is a way to control the time period of the system by inhibiting the rate of degradation HtlB has on CII in the oscillator. The protein used is the CIIICd, a DNA synthesised version of the CIIIC protein from the study done by Halder et al2.

The Reporter is the promoter PRE (the same part used in the oscillator) attached to a green flurorescent protein gene. It is used in various tests to check many of the components in the system.

References

  1. McMillen, D. et al (2002) "Synchronizing genetic relaxation oscillators by intercell signalling", Proc Natl Acad Sci USA, Vol. 22, No. 3, pp. 679-684.
  2. Hadler, S. et al (2007) Probing the Anitprotease Activity of λCIII, an Inhibitor of the Escherichia coli Metalloprotease HflB (FtsH)", Journal of Bacteriology, Vol. 189, No. 22, pp. 8130-8138.