Difference between revisions of "Part:BBa K325100"

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'''Description'''<br>
 
'''Description'''<br>
This part generates a mutant of the luciferase from the North American firefly (L.cruciata) as well as this species' luciferin regenerating enzyme (LRE). It is under the control of an Arabinose induced promoter (pBAD).  
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This part is a translational unit for a mutant of the luciferase from the North American firefly (P. pyralis) as well as this species' luciferin regenerating enzyme (LRE). It is under the control of an Arabinose induced promoter (pBAD).  
 
D-Luciferin has to be added to obtain light output.
 
D-Luciferin has to be added to obtain light output.
  

Revision as of 21:47, 23 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 part is a translational unit for a mutant of the luciferase from the North American firefly (P. pyralis) as well as this species' luciferin regenerating enzyme (LRE). It is under the control of an Arabinose induced promoter (pBAD). D-Luciferin has to be added to obtain light output.

The part is codon optimised for expression in E.coli.

THIS PAGE IS CURRENTLY BEING UPDATED.


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.