Generator

Part:BBa_K325219:ArabinosetoLight

Designed by: Bill Collins and Emily Knott   Group: iGEM10_Cambridge   (2010-09-15)
Revision as of 14:16, 24 September 2010 by PM (Talk | contribs) (→‎Data)

Input: L-Arabinose
Output: Light

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

Part Main Page        Arabinose -> Light        Add Data       


Description

This page describes the relationship between Arabinose concentration in the medium with light output. We used a [http://www.bmglabtech.com/products/microplate-reader/instruments.cfm?product_id=2 FLUOstar OPTIMA] microplate reader to quantify the light output. Protocol and plate reader settings used are given below.

Data

Figure 1 - Transfer function of BBa_F2620. The data points represent the mean of 6 or 9 individual measurements. The corresponding error bars represent the 95% confidence interval in the mean of the independent measurements. The blue shaded region represents the range bounded by the lowest and highest outputs among the independent measurements at a given input level. The solid black curve was calculated by fitting a simple Hill function to the experimental measurements.


Performance

Experiment1 Characteristic1 Value1
Transfer Function Maximum Output 6.6 PoPS cell-1
Hill coefficient 1.6
Switch Point 1.5E-9 M 3OC6HSL, exogenous
Response time: <1 min
Input compatibility Strong response to 3OC6HSL, C6HSL , C7HSL, 3OC8HSL, C8HSL
Weak response to C4HSL, C10HSL, C12HSL
Stability Genetic Stability
(Low/High Input) 		>92/>56 generations
Performance Stability
(Low/High Input) 		>92/>56 generations
Demand Internal Demand
(Low/High Input) 		Not measured
Transcriptional output demand:
(Low/High Input)
Nt = length of downstream transcript in nucleotides 		(0/6xNt) nucleotides cell-1 s-1
(0/1.5E-1xNt) RNAP cell-1
Growth Rate
(Low/High Input) 		54/59 min Doubling time

1Measured 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.