Difference between revisions of "Part:BBa K325219:ArabinosetoLight"

(Data)
(Protocol)
 
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==Data==
 
==Data==
  
[[Image:Arabinose_activation.png|thumb|569px|center|'''Figure 1 - Transfer function of <partinfo>K325219</partinfo>.  The data points represent the mean of 3 individual measurements. The corresponding error bars represent an interval of twice the standard deviation centred around the mean value. The values were obtained by integrating the light output obtained between 300 and 500 min after injection of D-Luciferin.''']]
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[[Image:IntegratedactiPP+LRE.png|thumb|569px|center|'''Figure 1 - Transfer function of <partinfo>K325219</partinfo>.  The data points represent the mean of 11 values obtained for light output at 30 min interval from 450 min to 750 min after injection of D-Luciferin. These values are the mean of 3 readings as shown in Figure 3. The corresponding error bars represent an interval of twice the standard deviation across the 33 data points centred around the mean value. ''']]
 
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[[Image:MaximumlumPP+LRE.png|thumb|569px|center|'''Figure 2 - Maximum luminescence output of  <partinfo>K325219</partinfo> as a function of Arabinose concentration. These values are the mean of 3 readings as shown in Figure 3. The corresponding error bars represent an interval of twice the standard deviation across the 3 data points centred around the mean value. ''']]
 
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[[Image:TimePP+LRE.png|thumb|569px|center|'''Figure 3 - Evolution of luminescence with time at different Arabinose concentrations. The interval between measurements is 30 min. Mean values and error bars are based on 3 time repeats.''']]
'''Performance'''<br>
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<center>
 
<center>
 
{|{{Table}}
 
{|{{Table}}
!Experiment<sup>1</sup>
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!Data
!Characteristic<sup>1</sup>
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!Notes
!Value<sup>1</sup>
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!Date Uploaded
 
|-
 
|-
|rowspan="3"|[[Part:BBa_F2620:Transfer Function|'''Transfer Function''']]
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|[[Media:BBa_K325219ArabinosetoLight.xls]]
|''Maximum Output''
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|Raw data from experiment
|6.6 [[PoPS]] cell<sup>-1</sup>
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|21/10/2010
|-
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|''Hill coefficient''
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|1.6
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|-
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|[[Switch Point|''Switch Point'']]
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|1.5E-9 M [[3OC6HSL|3OC<sub>6</sub>HSL]], exogenous
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|-
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|[[Part:BBa_F2620:Response time|'''Response time:''']]
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|<1 min
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|-
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|rowspan="2"|[[Part:BBa_F2620:Specificity|'''Input compatibility''']]
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|''Strong response to''
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|[[3OC6HSL|3OC<sub>6</sub>HSL]], C<sub>6</sub>HSL , C<sub>7</sub>HSL, 3OC<sub>8</sub>HSL, C<sub>8</sub>HSL
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|-
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|''Weak response to''
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|C<sub>4</sub>HSL, C<sub>10</sub>HSL, C<sub>12</sub>HSL
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|-
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|rowspan="2"|[[Part:BBa_F2620:Stability|'''Stability''']]
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|[[Genetic Stability|''Genetic Stability'']]<br>(Low/High Input)
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|>92/>56 generations
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|-
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|[[Performance Stability|''Performance Stability'']]<br>(Low/High Input)
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|>92/>56 generations
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|-
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|rowspan="4"|Demand
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|rowspan="1"|Internal Demand<br>(Low/High Input)
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|Not measured
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|-
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|rowspan="2"|[[Transcription Demand|''Transcriptional output demand:'']]<br>(Low/High Input)<br>Nt = length of downstream transcript in nucleotides
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|(0/6xNt) nucleotides cell<sup>-1</sup> s<sup>-1</sup>
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|-
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|(0/1.5E-1xNt) RNAP cell<sup>-1</sup>
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|-
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|[[Growth Rate|''Growth Rate'']]<br>(Low/High Input)
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|54/59 min Doubling time
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|}
 
|}
 
</center>
 
</center>
<sup>1</sup>Measured by the [http://2010.igem.org/Team:Cambridge Cambridge iGEM team 2010]
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 +
 
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</div>Measured by the [http://2010.igem.org/Team:Cambridge Cambridge iGEM team 2010]
 
<div style="padding: 00px; width: 680px">
 
<div style="padding: 00px; width: 680px">
 
'''Compatibility'''<br>
 
'''Compatibility'''<br>
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'''References'''<br>
 
'''References'''<br>
 
[http://www.ncbi.nlm.nih.gov/pubmed/18949818 '''[1&#x5d;:'''] 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.ncbi.nlm.nih.gov/pubmed/18949818 '''[1&#x5d;:'''] S.M. Marques and J.C.G. Esteves da Silva, (2009) Firefly Bioluminescence: A Mechanistic Approach of Luciferase Catalyzed Reactions,''Life'' '''61''', 6-17.
</div>
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 +
 
 
[http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html '''[2&#x5d;:'''] T. Nakatsu ''et al.'' (2006) Structural Basis for the spectral difference in luciferase bioluminescence, ''Nature'' '''440'''(16), 372-376.
 
[http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html '''[2&#x5d;:'''] 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&#x5d;:'''] 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.
 
[http://www.ncbi.nlm.nih.gov/pubmed/11457857 '''[3&#x5d;:'''] 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.

Latest revision as of 20:22, 27 October 2010

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_K325219. The data points represent the mean of 11 values obtained for light output at 30 min interval from 450 min to 750 min after injection of D-Luciferin. These values are the mean of 3 readings as shown in Figure 3. The corresponding error bars represent an interval of twice the standard deviation across the 33 data points centred around the mean value.
Figure 2 - Maximum luminescence output of BBa_K325219 as a function of Arabinose concentration. These values are the mean of 3 readings as shown in Figure 3. The corresponding error bars represent an interval of twice the standard deviation across the 3 data points centred around the mean value.
Figure 3 - Evolution of luminescence with time at different Arabinose concentrations. The interval between measurements is 30 min. Mean values and error bars are based on 3 time repeats.
Data Notes Date Uploaded
Media:BBa_K325219ArabinosetoLight.xls Raw data from experiment 21/10/2010


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.