Bioluminescence

Emily's Section

• Spectra:
  • Spectral Emissions of different species - Different Species Spectra
  • Fluorescent Protein Spectra - Spectra
  • Small amount of blue-shift - Blue Shift
  • More blue-shift - Blue Shift 2
• Metridia: (luciferase requires CBP Protein)
  • Metridia luciferase link - http://www.ncbi.nlm.nih.gov/pubmed/18754048
  • CBP link - http://www.ncbi.nlm.nih.gov/pubmed/18264586
  • CBP amino acid sequence http://www.rsc.org/ej/PP/2008/b713109g.pdf
• Firefly:
  • Oxyluciferin reduction - http://www.ncbi.nlm.nih.gov/pubmed/19872330
  • Luciferin Regenerating Enzyme (LRE) - LRE (in Photinus pyralis)
  • Firefly luciferin reaction - http://pubs.acs.org/doi/pdf/10.1021/ja00529a051
  • Light Emission Quanta - Spectral Emission and Quantum Yield of Firefly Bioluminescence
    • One light quantum emitted for every luciferin molecule oxidized
  • CoA stimulation - CoA in Luminescence
  • D-cysteine powerfully inhibits E.Coli growth D-cysteine
• Aequorin:
  • Regeneration of Aequorin in Aequorea Aequorea jellyfish - http://www.nature.com/nature/journal/v256/n5514/pdf/256236a0.pdf
• Bacteria:
  • Bacterial luciferase reaction - Bacterial Luciferin
  • Beetle luciferin - Putative Beetle Luciferin Synthesis
  • V. Fischeri Plasmid - V. Fischeri Plasmid
  • KCal/Mol luciferin in Lightning Bugs - Luciferin KCal/Mol
  • Photinus Pyralis (firefly) spectrum, 562nm - Reference
  • Different Coloured Firefly Luciferases - Colours
• Gaussia Princeps (copepod):
  • GLuc - GLuc
  • 100 Times Brighter than P. Pyrilus!! - So bright
  • GLuc expression in E. Coli E.GLuc

Theo's section

• Self contained part for Photorhabdus luminescens lux operon: http://partsregistry.org/Part:BBa_J70008:Design
• Firefly diagram from Emily's paper:

• Fireflow luciferin normally emits at 560nm but at higher temperatures at 630nm
• http://pubs.acs.org/doi/pdf/10.1021/bi00702a015 Kinetics
• Mg2+ required
• A nice background summary: http://irishmarinelife.com/bioluminescence-symbioses/
• The guy who made the biobrick's logbook: http://openwetware.org/wiki/User:Spenser_Skates/Notebook/pTKX_luciferase_biobrick/2008/04/14

Vibrio

• luxC = acyl coA reductase

Firefly

• The awesome paper http://www.springerlink.com/content/67w1q80435537747/fulltext.pdf

Biobricks related to light

Luciferases=

• Coelentrazine using
  • Renilla luciferase
• Firefly luciferin using
  • Firefly luciferase
• Produces all parts of pathway:
  • Lux operon from photorhabdus
• Click beetle:
  • Red

Fluorescent proteins

Bill's section

• Conversion of normal luciferin into firefly luciferin
• Don't use luciferinol, it r teh suckage
• Glowing fish what?
• Some actual numbers on luciferin [1]
• D-cysteine is bad for e-coli [2]

Will's section

• Kinetic analysis and modeling of firefly luciferase as a quantitative reporter gene in live mammalian cells.

• "Spectral emission and quantum yield of firefly bioluminescence" One rxn=>one photon

• General Notes for emission estimation

• Definition: Quantum Yield (Q)
• Definition: Katal

Anja's section

• Paper on mutant luciferase with increased activity
• Aequorin wavelength= 469 nm
• Low leakage rate
• Does not disrupt cell functions/ devt.
• Requires continual addition of luciferin
• Dr Laura Rowe's Paper on Bioluminescent Proteins
• Paper on Biosynthesis of Firefly D-Luciferin
• Paper on L-Tryptophan in Biosynthesis of Sea Firefly Luciferase

Peter's section

• Paper on various bacterial luciferase and luciferin synthesis pathways

Ben's section

• Flashlight fish symbiont luciferase genes have been sequenced but cannot be sucessfully expressed in E. Coli
• Suggested application http://www.yankodesign.com/2010/05/28/illuminate-your-influenza/
• The firefly luciferin regenerating enzyme is patented http://www.freepatentsonline.com/EP0825257.html
• Firefly luciferin biosynthesis appears not to be known. Vibrio Ficheri Lux operon is known. Would VF luciferin work with other luciferases? http://www3.interscience.wiley.com/cgi-bin/fulltext/89014035/PDFSTART

Paul's section

• Precursor to luciferin: http://www.sigmaaldrich.com/catalog/ProductDetail.do?lang=en&N4=261858|ALDRICH&N5=SEARCH_CONCAT_PNO|BRAND_KEY&F=SPEC
• Construction of a new firefly bioluminescence system using l-luciferin as substrate
• stuff to do with ATP in the cycle
• Applications of luciferin biosynthesis: Bioluminescence assays for l-cysteine and luciferase
• Biobrick for the Firefly luciferase for Slovenia '07: http://partsregistry.org/Part:BBa_I712019:Design
• Molecular cloning and expression of the cDNAs encoding luciferin-regenerating enzyme from Luciola cruciata and Luciola lateralis