Difference between revisions of "Part:BBa K325909"

 
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<div style="font-family: Arial; padding: 00px; width: 680px; border: 0px solid #000000;">
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{{Template:K325219 page header}}
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{{Template:K325219 Characterization Navbar}}
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<div style="padding: 00px; width: 680px; border: 0px solid #000000;">
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'''Description'''<br>
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This part generates a red-mutant of the luciferase from the Japanese firefly (L.cruciata) as well as the luciferin regenerating enzyme (LRE). It is under the control of an Arabinose induced promoter.
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D-Luciferin has to be added to obtain light output.
  
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__NOTOC__
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THIS PAGE IS CURRENTLY BEING UPDATED.
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<partinfo>BBa_K325909 short</partinfo>
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G28
 
  
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<!-- Add more about the biology of this part here
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'''Performance'''<br>
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===Usage and Biology===
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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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|-
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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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|}
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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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<div style="padding: 00px; width: 680px">
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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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<!-- -->
 
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<span class='h3bb'>Sequence and Features</span>
 
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<partinfo>BBa_K325909 SequenceAndFeatures</partinfo>
 
  
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</div>
  
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<!-- Uncomment this to enable Functional Parameter display
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'''References'''<br>
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===Functional Parameters===
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[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.
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<partinfo>BBa_K325909 parameters</partinfo>
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</div>
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<!-- -->
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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.
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[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.

Revision as of 10:57, 24 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 generates a red-mutant of the luciferase from the Japanese firefly (L.cruciata) as well as the luciferin regenerating enzyme (LRE). It is under the control of an Arabinose induced promoter. D-Luciferin has to be added to obtain light output.

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.