Team:NYMU-Taipei/Project/pLac

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The calibrated promoter has '''time-course standardized conversion''' between IPTG dosage and gene expression.  
The calibrated promoter has '''time-course standardized conversion''' between IPTG dosage and gene expression.  
Thus one can estimate the expression of unknown coding region ''X'' from calibration and compare with observed functional changes (e.g. the amount change of urea or phosphate to be removed, the period change of oscillators). This quantitative characterization procedure can precisely define the specification of functional genes between modeling and experimental verification.
Thus one can estimate the expression of unknown coding region ''X'' from calibration and compare with observed functional changes (e.g. the amount change of urea or phosphate to be removed, the period change of oscillators). This quantitative characterization procedure can precisely define the specification of functional genes between modeling and experimental verification.
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[[Image:QFC.png]]
[[Image:QFC.png]]
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* Rx (Rate of unknown gene expression)
* Rx (Rate of unknown gene expression)
* Rg (Rate of GFP expression in calibration system)
* Rg (Rate of GFP expression in calibration system)
* Rf (Rate of Functional change w.r.t. unknown functional gene X)
* Rf (Rate of Functional change w.r.t. unknown functional gene X)
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|}
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There are several technical issues should be concerned:
There are several technical issues should be concerned:

Revision as of 07:48, 30 October 2008

Contents

Methodology

It is an indirect method to quantitatively characterize the coding regions with specific functions (e.g. waste removal efficiency, tuner of oscillator) name QFCS (Quantitative Functional Calibration System). When we have an unknown coding region X, we can place this coding region after standard and calibrated Lac I promoter (pLac). The calibrated promoter has time-course standardized conversion between IPTG dosage and gene expression. Thus one can estimate the expression of unknown coding region X from calibration and compare with observed functional changes (e.g. the amount change of urea or phosphate to be removed, the period change of oscillators). This quantitative characterization procedure can precisely define the specification of functional genes between modeling and experimental verification.

QFC.png

  • Rx (Rate of unknown gene expression)
  • Rg (Rate of GFP expression in calibration system)
  • Rf (Rate of Functional change w.r.t. unknown functional gene X)


There are several technical issues should be concerned:

  • Rx =~ Rg (expression Rate of gene X can be estimated by GFP synthesis; However, it requires more detailed conversion, e.g. length of gene X, codon bias, etc.)
  • inconsistent growth between (pLac + GFP) and (pLac + gene X)

Calibration of pLac (R0010)

pLac + E0240 under 0 and 50uM IPTG

PLac 0 vs. 50uM IPTG.png

pLac + E0240 under 50, 500 and 1,000uM IPTG

PLac 50,500,1000uM IPTG.png

References

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