IGEM:MIT/2005/Notes

June 20, 2005

Divide & Conquer

• Sensor - External signal -> Internal Signal
  • Ray
  • Jessica

• Signal-Processor - Internal Signal Processing
  • Annie
  • Jenny

• Actuator - Internal Signal -> Output
  • Maxine
  • Will

• System Integration - What is this whole system doing?
  • Jen

• Read EVERYTHING by KT
  • Summarize relevant papers (Journal Club)
  • Don't make us look stupid.
• Please stock BUG Lounge with food. We are hungry college students
• We need to edit the Registry easier to read. More details.
• Plan assembly of test constructs
  • J07000: R0053, Q04121, Q04510, E0430
  • J07001: R0053, Q04121, Q04400, E0430
  • J07002: R0040, Q04121, Q04510, E0430
  • J07003: Z0251, Q04121, Q04510, E0430
  • J07004: Z0251, Q04121, Q04400, E0420
• Place order with Registry for parts to be synthesized
  • As said in above bullet.
  • We need R0010. Has CAP binding site.
• Prepare FACS samples for Thursday
  • We will get cells from Drew's Lab.
• Order new parts from Blue Heron
  • Specify on Registry!
    • insulin - Karen Talmadge
    • export tag
    • etc?
• Order supplies
• Make stocks

Test Constructs

How are we going to test our system in its intermediate, and also its final stages?

1. Flow cytometry
2. Beta gal assay
3. Microscopy (detect gfp, bgal, etc)
4. Western blot (measure protein levels)
5. insulin assay?
6. use cells as indicator - as cell-cell signalling system!
   • insulin receptor (research this)
   • sender/receiver
   • feedback (incorporate insulin output into regulating glucose - ultimately CAP - input)
7. RT-PCR
   • used to measure RNA levels
8. Northern Blot
   • used to measure RNA levels

June 15, 2005

• Familarize
  • Glucose receptor (what is the signal cascade?)
    • plasmid?
    • chromosome?
  • Insulin plasmid (read the paper about it)
  • How is insulin produce from E. coli?
• Tags
• Sequence of insulin
• Apoptosis in E. coli?
• List of things to get for lab
• Insulin is in 2 chains, so we need to look at how we can regulate it. In vitro?