User:Maira Tariq/sandbox: Difference between revisions

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*[[Image:ICGEMS_dvc_PHrpL_x-gal.PNG‎|left|thumb|200px|]]
*[[Image:ICGEMS_dvc_PHrpL_x-gal.PNG‎|left|thumb|200px|]]
*[[Image:ICGEMS_sys_Ptet_luxR_Plux_x-gal.PNG‎|left|thumb|400px|]]
*[[Image:ICGEMS_sys_Ptet_luxR_Plux_x-gal.PNG‎|left|thumb|400px|]]
==[[IGEM:IMPERIAL/2007/Projects/Experimental_Design/Problems|Problems page]]==
*Contains a list of all the problems being faced by the team regarding the experiments being carried out

Revision as of 13:41, 5 September 2007

Application driven projects: (23/07/07)

  • Detecting food spoiling
    • Detect pH change and temp change in surrounding environment
    • Visual change to show spoiling
  1. Ternström A, Lindberg AM, and Molin G. Classification of the spoilage flora of raw and pasteurized bovine milk, with special reference to Pseudomonas and Bacillus. J Appl Bacteriol. 1993 Jul;75(1):25-34. DOI:10.1111/j.1365-2672.1993.tb03403.x | PubMed ID:8365951 | HubMed [Martino]
  1. can use bacteria to detect biofilm on surface.
  2. don't introduce bacteria inside as may accumulate
  3. discard the catheter if biofilm detected
  • Detect food for allergic substances eg nuts, dairy products etc.
  • Indicate excess UV exposure of skin
  • Biofilm to form lubricant(cartilage) on joints
    • Biocompatible
  • Stimulate bacteria in a contolled way
    • Create 3D images
    • Biological-electronic interface
  • Gut bacteria - assess nutritients in diet
  • Layer of biofilm on stagnant water to block sunlight and oxygen
    • Kill mosquito cells
  • HRP system applications:
    • Glucose/insulin regulation system
    • Problem: no receptor in E. coli to detect glucose directly

AHL produced in biofilms:

(Notes on the journal modeling AHL production in biofilms of the same bacterial population) AHL synthesis is subject to autoinduction in which production of AHLs operates as a positive feedback loop.
Assumptions made in the model:

  • All bacterial cells are physiologically identical with regard to size, shape and permeability of the cell membrane, as well as production and degradation rates of the signalling molecules
  • Bacterial population exhibit a standard logistic growth pattern
  • No metabolic or physiological lag is assumed
  • At very low Cbc, the net rate of AHL production, h(Cbc), is assumed to be determined solely by the difference between basal production, Bp, and degradation of AHLs
  • Degradation of AHLs is proportional to the concentration of AHL and occurs at a rate d*Cbc

Not considered in the model: permeability constant a, which is characteristic of the bacterial cell membrane, the diffusability of a given AHL, and the viscosity of the cell and the biofilm

Conclusions from the model: high concentrations of AHL inside cells could be achieved at very low population densities. Rapid rise in AHL concentration early in population growth, followed by a plateau, followed by another rise to a second plateau

Biofilm detection using AHL as signals

http://aem.asm.org/cgi/reprint/67/2/575

Cell-free Schedule

Week 3 Week 4 Week 5 Week 6 Week 7 Week 8
Design EBF
Model ECBD
Protocol EHRP
Experiment:
EBF I
ECBD I
Making EHRP Testing EHRP
Test-tube/In-vitro Characterization Design/Mod
TT BF
TT CBD
Expt/Test
TT BF I/II/III
TT CBD I/II/III
In-Veso: Characterization Mod/Design
VBF I/II/II
VCBD I/II/III
Expt/Test
VBF I/II/II
VCBD I/II/III

Colour Scheme

Orange indicating E-Coli

Red indicating "Pure" cell-free extract

Green indicating Vesicular form



Constructs


  • Ptet promoting LuxR, Plux promoting HrpR and HrpS
  • PHrpL promoting GFP

Problems page

  • Contains a list of all the problems being faced by the team regarding the experiments being carried out