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Team:NYMU-Taipei/Project/pLac
From 2008.igem.org
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== Methodology == | == Methodology == | ||
| - | It is an indirect method to quantitatively characterize the coding regions with specific functions | + | It is an indirect method to quantitatively characterize the coding regions with specific functions (e.g. waste removal efficiency, tuner of oscillator) |
| + | 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) | ||
== Calibration of pLac (R0010) == | == Calibration of pLac (R0010) == | ||
Revision as of 05:50, 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) 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)
Calibration of pLac (R0010)
pLac + E0240 under 0 and 50uM IPTG
pLac + E0240 under 50, 500 and 1,000uM IPTG
References
- [http://science.nhmccd.edu/biol/operon.html gene regulation and operon]
- [http://rothlab.ucdavis.edu/protocols/beta-galactosidase-3.html b-Galactosidase Activity Assay]
- [http://www.bio.davidson.edu/Courses/Molbio/MolStudents/spring2003/Causey/pET.html The pET Expression System]
- [http://www.escience.ws/b572/L11/L11.htm The lac operon]
- [http://www.springerlink.com/content/8texkfla66v0hcxc/ The Effect of the lacY Gene on the Induction of IPTG Inducible Promoters, Studied in Escherichia coli and Pseudomonas fluorescens, Current Microbiology 1997]
- [http://www.pnas.org/content/104/14/6043.abstract Combinatorial transcriptional control of the lactose operon of Escherichia coli, PNAS 2007 (Thomas Kuhlman and et. al)]
