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Bioluminescence

Uses and application in the real world. Bioluminescence. Uses. Bioluminescence Resonance Energy Transfer (BRET) ‏ Bioluminescent molecules in gene expression Identification of bacterial contamination in food Artistic inspiration. BRET: Bioluminescence Resonance Energy Transfer.

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Bioluminescence

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  1. Uses and application in the real world Bioluminescence

  2. Uses Bioluminescence Resonance Energy Transfer (BRET)‏ Bioluminescent molecules in gene expression Identification of bacterial contamination in food Artistic inspiration

  3. BRET: Bioluminescence Resonance Energy Transfer BRET is a proximity-based assay where the energy generated by the catalytic degradation of coelenterazine by the enzyme Renilla luciferase (Rluc) (energy donor) is transferred to a green fluorescent protein (GFP) acting as the energy acceptor. The GFP then emits light at its specific emission wavelength.

  4. Degradation of coelenterazine by the enzyme Rluc

  5. BRET

  6. BRET: Bioluminescence Resonance Energy Transfer BRET can be used to observe protein-protein interaction in living mammalian cells. It is based on the non-radioactive transfer of energy between a luminescent donor Rluc and a fluorescent acceptor (Green Fluorescent Protein or GFP). May be used in the future to identify new protein complexes in human.

  7. Monitoring of ubiquitination in living cells by BRET Ubiquitination is descried as the rapid process of post-translational modification present in many aspects of biology involving a covalent attachment of ubiquitin to proteins.

  8. Monitoring of ubiquitination in ling cells with BRET Upon degradation of DeepBlueC,the Rluc fuses to β-arrestin and emits blue light with an emission peak at 395 nm. If the β-arrestin is ubiquitinated, a transfer of energy will occur between Rluc and the GFP2 fused to the N terminus of ubiquitin, resulting in re-emission of light with an emission peak at 510 nm.

  9. Continuous light spectrum

  10. Gene expression Bioluminescence can be used to study prokaryotic gene expression inside living cells It allows the observation of biological processes in real time, as they happen. This technique can be used as a noninvasive way to study protein trafficking, protein function, genetic regulatory or image bacteria, tumors and genes over a long period time.

  11. Applications of bioluminescent imaging. (a) assessing the levels of trans-gene expression, (b) the location and extent of bacterial infection, (c) the efficiency of gene transfer and expression, and (d) the trafficking patterns of lymphocytes.

  12. Bioluminescence in gene expression Luciferases are light-generating enzymes that can be found in bacteria, marine crustaceans, fish and insects. Luciferases are nontoxic and can be injected to become gene expression markers.

  13. Gene Expression in Yeast Cells Cells fused to GFP, making the component protein of microtubules (alpha-tubulin)‏ The alpha-tubulin:GFP fusion can be observed by exciting the GFP, causing a green light emission. It can be used to study gene coding mutations such as kinases.

  14. Bioluminescence in Food Industry Methods of use ATP bioluminescence Bacterial bioluminescence

  15. ATP Bioluminescence Measures the amount of ATP that is converted to photons of light by living cells. The amount of light emitted to proportional to the number of bacteria in a food sample It can be used to measure the number of lactic acid bacteria in a contaminated food sample. Firefly luciferase is used to detect the presence of ATP

  16. Luciferin and the reaction with ATP

  17. Bacterial Bioluminescence Can be used to test for a specific bacterial species or possible food born pathogen such as Salmonella typhimurium. The lux gene, responsible for bacterial bioluminescence, has been isolated and cloned. The cloned lux gene can be injected into a host-specific phage, which does not have the ability to express the gene. If the phage infects a host bacterium, light emission results.

  18. Detection of Bacteria

  19. Artistic Inspirations Poetry Photography Painting

  20. Fireflies, By Edgar Fawcett I saw, one sultry night above a swamp, The darkness throbbing with their golden pomp And long my dazzled sight did they entrance With the weird chaos of their dizzy dance! Quicker than yellow leaves, when gales despoil, Quivered the brilliance of their mute turmoil, Within whose light was intricately blent Perpetual rise, perpetual descent. As though their scintillant flickerings had met In the vague meshes of some airy net! And now mysteriously I seemed to guess, While watching their tumultuous loveliness, What fervor of deep passion strangely thrives In the warm richness of those tragic lives, Whose wings can never tremble but they show Those hearts of living fire that beat below!

  21. Still Photography Artistic re-creation of the Bioluminescent Bay in Vieques

  22. Sculpture Montana State University-Bozeman Bioglyphs project Collaboration between art and science during 2002 by members of the center for Biofilm Engineering and the MSU School of art

  23. Lights on

  24. Lights off

  25. Lights on

  26. Lights off

  27. Involves the practice of "painting" on prepared Petri dishes with a sort of "invisible ink" composed of liquid medium inoculated with the bacteria. The microorganisms themselves went to work, multiplying on the plates and beginning to produce light within 24 hours. The only light available to view the art was that produced by the bacteria themselves. Over the five-day period, the light intensity of the paintings changed as the bacteria multiplied and then gradually consumed the nutrient available. Bioglyphs

  28. Gallery, with lights on

  29. Gallery, with lights off

  30. Sources Stephane Angers, Ali Salahpour, Eric Joly, Sandrine Hilairet, Dan Chelsky, Michael Dennis, and Michel Bouvier. “Detection of β 2-adrenergic receptor dimerization in living cells using bioluminescence resonance energy transfer (BRET).” Proceedings of the National Academy of Sciences 97(2000): 3684-3689. http://www.pnas.org/cgi/content/full/97/7/3684 (5 October 2007)‏ Claire Normand1, Stéphane Parent1, Benoit Houle, Anne Labonté, Lucie Bertrand, Mireille Caron, Mireille Legault, Stéphane Angers, Michel Bouvier, Erik C. Joly and Luc Ménard. “BRET2™: Bioluminescence Resonance Energy Transfer, a Novel Assay Technology to Examine GProtein Coupled Receptor Activation in Intact Cells.” 2002 http://las.perkinelmer.com/Content/RelatedMaterials/Posters/PSH_BRET2NovelAssayTechnology.pdf (9 October 2007).

  31. Saman Hosseinkhani. “Bioluminescnece.” 2007 http://www.modares.ac.ir/sci/saman_h/Pages/Bioluminescence%20.htm (7 October 2007). M. W. Griffiths. “Applications of Bioluminescence in the dairy Industry.” Journal of Dairy Science 76 (1993): 3118-3125. http://jds.fass.org/cgi/reprint/76/10/3118.pdf (2 October 2007). Julie Perroy, Stephanie Pontier, Pascale G Charest, Muriel Aubry and Michel Bouvier. “Real-time monitoring of ubiquitination in living cells by BRET.” 1 (2004): 203-208. http://www.nature.com/nmeth/journal/v1/n3/abs/nmeth722.html;jsessionid=A04B9606CE4D3B8FB5147E17BDEF094D (6 October 2007). Bouvier;s Lab. “Using BRET to study Protein-Protein interactions.” June 5, 2005. http://www.mapageweb.umontreal.ca/bouvier/bret/index.html (8 October 2007). Montana State University. “Bioglyphs.” January 3, 2007. http://www.erc.montana.edu/Bioglyphs/default.htm (9 October 2007). Jackie Vogel. “Research.” June 3, 2005. http://biology.mcgill.ca/faculty/vogel/research.html (10 October 2007)‏

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