Presentation

From 2006.igem.org

The Powerpoint can be found here: [http://www.individual.utoronto.ca/chuck_1985/UT.Waterloo.iGEM.2006.Presentation.ppt Presentation]

University_of_Toronto_2006

Presentation Script

Slide 1

• Introduce the team (i.e. collaborative effort between UofT and UW) and mention the project we're working on.

Slide 2

• Explain what makes this thermometer unique from all other existing models as in other models may limitations to where/how they can be used
  • size of the species of interest may be too small
  • research experiments where chemicals, radiation may be toxic for a colony but temperature readings are needed

• A bio-synthetic thermometer may be a possible solution to overcome these obstacles.

• Applications:
  • use to detect small changes in temperature in very small organisms- we can do this by measuring the intensity of the colour emitted
  • experiments or places where a quantitative reading cannot be obtained (i.e a person can not physically be there to take the reading (in anaerobic conditions?)) or where air pressure
  • may also use this to monitor temperature of aquariums using zebra fish for example

Slide 3

• Explain diagrams

Slide 4

• Explain our approach: how we decided to ligate parts separately and build up to the final product
• Constructed many new parts
• Replaced our cI repressor system with tetracycline repressor (tetR), an analogous system to cI but less leaky and can control externally

Slide 5

• Testing Phase
  • Intro: - Testing was split into 3 modules.
    • Each module acts as a checkpoint for a particular milestone in the construction process.
    • After each module we can ensure a vital component of the construct is functional.
  • State the goal/purpose of each module and how it was executed.

Slide 6

• Test Results
  • Tested the intermediary pBad/araC + LacI ts coding sequence (R0011) + reporter gene thoroughly
  • We ensured that module 1 tests passed before continuing construction.

Slide 7

• Conclusions/Results!
  • Does it work/or not?
  • What we learned from constructing the device
    • at the beginning, we tested all enzymes for functionality via test plasmid + running a gel - while doing this we learned that Xba1 works slower than EcoRI, SpeI, and PstI
    • Team communication, allocate more time for testing in addition to construction/designate a test team
  • Limitations to the device
    • can only detect temperatures in a small range
    • does fluorescent protein stay? or decay quickly? ask Charles/Andy
    • Experiments involving radiation - this thermometer cannot be used as the radiation will damage the DNA
    • Expression of the reporter gene must be as sensitive as the activation of the transcription factor (for accuracy in timing events dependent on temperature and time)
    • Future work that can be done to improve the device/design for next year
    • investigate repressors etc that would be good for larger temperature ranges
    • this would increase the application of the device allowing it to be another alternative to detecting temperature changes in biological systems.

Slide 8

• Team Members

Slide 9

• Acknowledgements