IGEM:IMPERIAL/2007/Projects/Hrp System

HRP Characterization

Things to Characterize

What we want to know (in order of importance):

1. Strength of response
2. Minimum/maximum PoPs output
3. Signal fluctuation and noise
4. Leakiness
5. Response times
6. Stability of proteins (half life)
   • Recovery of response
7. Ratio effects on output
8. Interaction with other parts

• How R and S affect activity separately
  • Having both R and S under same promoter
• Energy requirements
• Enhancer length effects

We still need to:

• Find literature for each question
• Design experiments for each question
  • Note down the time requirements
• Plan the schedule

Measuring PoPs

PoPs depends on:

1. Temperature
2. Medium
3. pH

• IHF
• PspF
• LoN protease
• Other regulators like σ₅₄
• Host cell

Other things that affect PoPs:

• Promoter
• Enhancer length

How do we measure PoPs? Can we do it using the screening plasmid?

Contexts

Measuring transfer function in E.coli

1. Normal genome E.coli in vivo
2. in vitro
3. E.coli with other parts
4. Minimal genome E.coli in vivo