IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Notes: Difference between revisions

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=Biofilm Detection: Notes=
=Infector Detector: Notes=
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<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Design|Design]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Design|Design]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Modelling|Modelling]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Modelling|Modelling]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Implementation|Implementation]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Implementation|Fabrication]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/TestingValidation|Testing/Validation]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/TestingValidation|Testing]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Data Analysis|Data Analysis]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Validation|Validation]]</li>
<li id="current">[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Notes|Notes]]</li>
<li id="current">[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/Notes|Notes]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/References|References]]</li>
<li>[[IGEM:IMPERIAL/2007/Projects/Biofilm Detector/References|References]]</li>
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==Making Biofilm==
*Freshly-grown growth-phase E. coli ZK1056 culture is added to sterile medium to a concentration of 5 × 107 cells/mL.
*Round 15 mm sterile glass coverslips are placed upright in the wells of a 24-well plate with sterile plastic stands
*Add 1 mL of E. coli solution to each well.
*The 24-well plate is immediately covered and transferred to a 30 °C incubator.


==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.
<br> 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


==Project Schedule==
==Biofilm detection using AHL as signals==
===Week 3 - 4===
http://aem.asm.org/cgi/reprint/67/2/575
'''Specific Reading:'''
*Diffusion of vesicles
*Concentration of AHL in biofilms
*Alternatives amplifiers to HRP
 
'''Modelling of system'''
 
 
Background Reading:
*General information on biofilms
*Further reading on application of system
*Suitable gels for diffusion
*Catalog search for AHL
 
What to do by end of Week 4 Wednesday:
*Write proper specifications
*Protocols of experiments on specific parts
 
===Week 4 - 5===
*Building basic constructs
*Testing basic constructs in E.coli
*Plan experiments with HRP and cell free system
 
===Week 6 - 7===
*Build constructs with HRP and luciferase
*Test in cell free system

Latest revision as of 07:55, 26 September 2007

Infector Detector: Notes



Making Biofilm

  • Freshly-grown growth-phase E. coli ZK1056 culture is added to sterile medium to a concentration of 5 × 107 cells/mL.
  • Round 15 mm sterile glass coverslips are placed upright in the wells of a 24-well plate with sterile plastic stands
  • Add 1 mL of E. coli solution to each well.
  • The 24-well plate is immediately covered and transferred to a 30 °C incubator.

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