Difference between revisions of "Part:BBa K325100"

 
 
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<partinfo>BBa_K325100 short</partinfo>
 
<partinfo>BBa_K325100 short</partinfo>
  
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This is a mutant luciferase that is very much brighter than wild-type. It is supplied in an operon with an enzyme (LRE) that regenerates its substrate. When supplied with luciferin and placed under a promoter it will emit light at a green wavelength. If D-cysteine is added the LRE will be able to regenerate the luciferin substrate.
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'''Description'''<br>
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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).  
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D-Luciferin has to be added to obtain light output.
  
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<!-- Add more about the biology of this part here
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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.
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===Usage and Biology===
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The part is codon optimised for expression in ''E.coli''.
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K325100 SequenceAndFeatures</partinfo>
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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_K325100 parameters</partinfo>
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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.

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.