IGEM:UNAM Genomics Mexico/2009/Notebook/Wifi coli/2010/05/04

Modeling & WetLab Meetings

Modeling

Stoichiometry of Luciferase activity.

a[Luc] + b[ Luc-ina] + c[02] + d[ATP] -> z[Luc] + y [oxyLuc-ina] + w[C02] + u[AMP] + v[PPi]

We use that formula to know which is the limitant reactant and the optimum concentration.

The next figure is to show the posible conectreations of Luciferin and Luciferase.

To achieve a controlled model we use single copy plasmid or low copy number.

LuM= Kc_cat * LUC * (Scyt/(Km+Scyt)) *RLV

K_cat= [Constant of Activity]

LUC= [Luciferase]

Scyt = [Cytosolic Luciferase]

Km= Michaelis-Menten constant.

RLU= Relative Light Unit

LuM= Number of moles, which react in measure, this is by colony.

With this formula we can achieve the values for light emission and efficiency of our system. We need:

• Get values of K_cat.
• Complete equation to make more robust and include more parameters with degradation and transcriptional activity.
• Take note about the double action of Luciferase, Luminescence and quiral activities.

To get unidirectionality we purpose a system with optical fiber and aisolated systems.

Each beaker is coated by aluminium to focus photons to exit. This design is simple, we will change datails in sizes of elements and positions.

Wet lab

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