IGEM:Cambridge/2008/PresentationOutline

Voltage

• KDP
  • K+ ATPase, import pump
  • PCRed from genome
  • OsmY

• Flame Photometry
  • Data and graphs

• GluR0
  • Glutamate gated K+ channel
  • From Synechosystis
  • Constitutive Promoter

• Voltage Output
  • Pictures

Peptide Signaling

• AGR System
  • Sender & Reciever

• Compared to Lux
  • More robust, specific
  • Controlled secretion, not membrane diffusion
  • Multichannel signaling (no crosstalk)
  • Requires expression of membrane channels

Bacillus

• About Bacillus
  • Pretty Pictures
  • Best-characterized Gram Positive
  • Very safe
  • Genome Sequenced
  • Protein Secretion
  • Not compatible with Gram-neg promoters/RBS

Techniques/Tools:

• Chromosomal Integration
  • Single Copy
  • Cool diagram: single vs. double x-over
  • Detection: amylase assay & PCR
• InFusion
• New Vectors
• Bacillus RBS & Promoters
  • Two Constituitive, Two Inducible
  • Culled from existing vectors
  • Picture of Bacillus expressing GFP with pUpp
• New GFP
  • Stefan's page

Modeling

The agr-peptide signalling system.

Assumptions.

Results: Step-like behaviour, system highly sensitive to basal rates of expression.

Pattern formation.

Some simple models.

Patterns with agr-quorum sensing systems: competing populations.

Further questions: Discrete models, spatial pattern generation with a single signalling molecule.