# IGEM:UNAM Genomics Mexico/2009/Notebook/Wifi coli/2010/05/25

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## Represilator

### A brief review to compare

• What?.

A loop negative system.

• Parameters?.

Degradation of the mRNA, protein and the rate of transcription.

### Perspectives for our system

• Goes to a stable state.

Limitants -> lifespan of E. coli?, luciferin?.

"If is all on, we expect, that stuck there, but if one system turn off, maybe there is a stochastic component".

## LOVTAP System

### Lifespan TetR

• We need a tag for increase the degradation of our repressor.

$\displaystyle δTetR= β - αTetRο$

We require a formula:

$\displaystyle F([TetR]) = Pops$

But what happen with the production of endougenos TetR?, exists?.

• No... haha, it seems to be in a RESISTANCE plasmid.

## Green emission

Input Pops -> Photons.

τ → ∞ -> $\displaystyle [14-CHO]=k$

• This because the action of deg. and the production of CDE by M. acid tend to stabilize

Again we need to propose a formula:

$\displaystyle [LuxAB]=Pops=N*f(Promoter force)$

### Pathway

$\displaystyle LuxAB-FMNH$ 2$\displaystyle -O$ 2$\displaystyle -R-CHO -> LuxAB + FMN + R-COOH + H$ 2$\displaystyle O + Light$

And the enzymes work in this form (Daniela has the references in German :S):

$\displaystyle Acid M. -$ LuxD$\displaystyle > C$ 13$\displaystyle H$ 27$\displaystyle -COOH -$ LuxE$\displaystyle >C$ 13$\displaystyle H$ 27$\displaystyle Co-Amp -$ LuxC$\displaystyle > 14-CHO$

Here we see:

• It appears that the product of the aldehyde can enter in the recicle pathway of LuxC and LuxE
• We assume that C & E enzymes will be ocuppied with more substrate than the D, that only receives the membrane acid to form R-COOH. This mean, we need a major concentration of LuxCE than LuxD

### Perspectives

It is relevant?:

• Formation of the dimer A|B
• Formation of the complex FMN-AB
• Formation of the complex R-CHO-FMN-AB
• Temperature

Can we assume Micahel-Menten equation?

## To find list

• Rate of deg. 14-CHO
• Distribution of the acid trough the membrane
• Size of the cell
• Rates of transcription
• Kcat´s and Km´s
• Parameters that affects the emission.