Team:Rutgers/BEAS

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Rutgers 2012 iGEM Team: Biofuels in Biology

Rutgers 2012 iGEM Team: Biofuels in Bacteria

Abstract

The Etch-a-Sketch project aims to create a lawn of bacteria that can be drawn on with a laser pointer. There are quite a few obstacles in building this circuit. For example, the bacteria need to be sensitive enough to respond to a short light pulse from a laser, but still “selective” to not respond to ambient lighting.

We have designed a novel genetic switch to tackle these problems. This project will serve as a vital step to any other project requiring amplification. For example, researchers creating biosensors may find our work very helpful.

 

See our project from last year and read our current progress and results.

Plasmid Maps

High copy and low copy plasmid maps

A new simplified first run target. This is our general plan for the circuit. If this runs, the bacteria will express mRFP and thus respond to the selected wavelength of light.

 

Plasmid 1

  • Plasmid 1 ligation plan was completed.
  • This plasmid is carrying the LovTap protein which activated the circuit due to it's sensitivity to blue light (470 nm)
  • mRFP in this plasmid is used as a sensor and an assay for the circuit.

 

Plasmid 2

  • Plasmid 2 is carrying most of the genes involved in the forming the locking switch which enables the bacteria to become independent from the light source.
  • This plasmid was completed excluding the last ligation (L23)
  • There needs to be a double terminator which will be inserted via roundhorn mutagenesis.

 

 

Results and Sucessess

pTrpL and pSB1C3 gel results

Here we show our first sucessful cloning of

  • pTrpL - which was designed as oligos
  • using EcoRI and PstI on the plasmid pSB1C3

 

Read more from our lab notebook

Read more about the Etch-a-Sketch circuit from RUiGEM2011