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. Response times
5. Stability of proteins (half life)
   • Recovery of response
6. Leakiness
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

Measuring Pops

Pops depends on:

• Promoter
• Enhancer length
• Temperature
• pH
• Medium

Other physical sensitivities include:

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

Contexts

Measuring transfer function in E.coli

• in vitro
• in vivo
  • Minimal/Normal medium E.coli
  • E.coli at optimal temperatures
  • E.coli with other parts