Difference between revisions of "Part:BBa K325100"
 
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<partinfo>BBa_K325100 short</partinfo>
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{{Template:K325219 page header}}
 
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{{Template:K325219 Characterization Navbar}}
 
 
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'''Description'''<br>
 
'''Description'''<br>
 
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).  
 
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.
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EPIC stands for '''Enhanced Photon Initiating Complex'''. It is described in [http://www.ncbi.nlm.nih.gov/pubmed/17540326 Fujii et al. 2007] as having a 10 times higher substrate affinity and luminescence output compared to wildtype.
  
 
The part is codon optimised for expression in ''E.coli''.
 
The part is codon optimised for expression in ''E.coli''.
  
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THIS PAGE IS CURRENTLY BEING UPDATED.
 
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'''Performance'''<br>
 
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<center>
 
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{|{{Table}}
 
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!Experiment<sup>1</sup>
 
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!Characteristic<sup>1</sup>
 
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!Value<sup>1</sup>
 
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|-
 
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|rowspan="3"|[[Part:BBa_F2620:Transfer Function|'''Transfer Function''']]
 
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|''Maximum Output''
 
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|6.6 [[PoPS]] cell<sup>-1</sup>
 
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|''Hill coefficient''
 
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|1.6
 
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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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|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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|[[Performance Stability|''Performance Stability'']]<br>(Low/High Input)
 
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|>92/>56 generations
 
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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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|(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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|}
 
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</center>
 
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<sup>1</sup>Measured by the [http://2010.igem.org/Team:Cambridge Cambridge iGEM team 2010]
 
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'''Compatibility'''<br>
 
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[https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=cell ''Chassis:''] Device has been shown to work in ''Top 10 (Invitrogen)''<br>
 
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[[Plasmid backbones|''Plasmids:'']] Device has been shown to work on ''<partinfo>pSB1C3</partinfo>'' <br>
 
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</div>
 
  
 
'''References'''<br>
 
'''References'''<br>

Latest revision as of 03:49, 28 October 2010

EPIC Firefly Luciferase and LRE
P. Pyralis
(E. coli optimised)

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.

EPIC stands for Enhanced Photon Initiating Complex. It is described in [http://www.ncbi.nlm.nih.gov/pubmed/17540326 Fujii et al. 2007] as having a 10 times higher substrate affinity and luminescence output compared to wildtype.

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


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.