Generator

Part:BBa_K325219:Stability

Designed by: Bill Collins and Emily Knott   Group: iGEM10_Cambridge   (2010-09-15)
Input: L-Arabinose
Output: Light

pBad/araC
I0500		Luciferase/LRE
K325210
 Cambridge-Eglowli.png

Part Main Page        Arabinose -> Light        Add Data       


Description

Both the intensity and the spectrum emitted by the luciferase-luciferin reaction has been shown to be higly dependent on the pH of the medium. The main characterisation experiments have been performed in LB Broth at pH 7, so in order to assess this effect cultures with LB and a citrate buffer were prepared (pH = 5.3, pH 6.1 and pH = 7) This page describes the results of these experiments. We used a [http://www.bmglabtech.com/products/microplate-reader/instruments.cfm?product_id=2 FLUOstar OPTIMA] microplate reader to quantify the light output. Protocols and plate reader settings used are given below.

Data

Figure 1 - Maximum light output within 5 hours of D-luciferin injection at different pH values. These values are the mean of 3 readings. The corresponding error bars represent an interval of twice the standard deviation across the 3 data points centred around the mean value.
Figure 2 - Evolution of light output at different values of pH. Measurements are taken every 20 min. These values are the mean of 3 readings. The corresponding error bars represent an interval of twice the standard deviation across the 3 data points centred around the mean value.
Data Notes Date Uploaded
Media:BBa_K325219pheffect.xls Raw data from experiment 21/10/2010

Protocol

  1. The protocol can be found as [http://2010.igem.org/Team:Cambridge/Notebook/Week11 Experiment 110] on the [http://2010.igem.org/Team:Cambridge Cambridge iGEM 2010 Website].


Measured by the [http://2010.igem.org/Team:Cambridge Cambridge iGEM team 2010]

Compatibility
Chassis: Device has been shown to work in Top 10 (Invitrogen)
Plasmids: Device has been shown to work on pSB1C3


References
[http://www.ncbi.nlm.nih.gov/pubmed/18949818 [1]:] S.M. Marques and J.C.G. Esteves da Silva, (2009) Firefly Bioluminescence: A Mechanistic Approach of Luciferase Catalyzed Reactions,Life 61, 6-17.


[http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html [2]:] T. Nakatsu et al. (2006) Structural Basis for the spectral difference in luciferase bioluminescence, Nature 440(16), 372-376.


[http://www.ncbi.nlm.nih.gov/pubmed/11457857 [3]:] K. Gomi and N. Kajiyama, (2001) Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin, The Journal of Biological Chemistry, 276(39), 36508-36513.