User:Daniela A. Garcia S./Notebook/Modeling UNAM-Genomics Mexico/2010/05/18

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Modeling Meeting: Luciferin Photinus pyralis System

  • The change in the luciferin concentration is up to three parameters: reaction, degradation and LRE.
    • The degradation function makes used of the constant of degradation for the luciferin.
    • The reaction function needs the concentration of the luciferin and the luciferase. All in an Michaelis-Menten ecuation.
    • For the LRE reaction we proposed a Michaelis-Mented nested. At the end due to the stochasticity of the process (D- and L-luciferin) we assumed an effective concentration of one half.

General

From the article: "Kinetic Analysis and Modeling of Firefly Luciferase as a Quantitative Reporter Gene in Live Mammalian Cells"

  • 100 uM of D-luciferin in water was added
  • Km lower than the one for live cells
  • High luciferase concentrations inside the cells. Localization of the enzyme inside the cells unable to access the free luciferin in the surrounding media.
  • Initial level of luciferase, standard curve based on purified recombinant liciferase from Promega.
  • Luciferin consuption approximately 0.5nM/s
  • Gradual increase in the value of the signal due to cell proliferation


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