An Illuminating Presentation

2. Dave Quiles Imagineering Productions Presents An Illuminating Presentation In Illumicolor (Where Available)

3. Davey Q Flash Firefly

4. Alrighty, what exactly is bioluminescence? Bioluminescence is a natural phenomenon in which organisms produce and emit light via an enzyme-mediated biochemical process. This visible light is ìcoldî- meaning that, unlike incandescent sources, light generated by bioluminescence does not give off much energy in the form of heat.

5. Selected History of Bioluminescence • 500 B.C. Aniximenes • He described light exhibited by the sea when struck • with an oar. • 350 B.C. Aristotle • Considered lightning similar to when "you strike the sea with a rod by night and the water is seen to shine". • 1637 A.D. Rene Descartes • Striking seawater will "generate • sparks rather similar to those which • are emitted by pieces of flint when • they are struck."

6. Selected History of Bioluminescence, Continued • 1668 A.D. Robert Boyle • ìI am tempted to supect, that some cosmical law • or customof the terrestrial globe, or, at least, of • the planetary vortex, may have a considerable • agency in the production of these effects.î • 1717 A.D. Sir Isaac Newton • "...do not all Bodies...emit Light as • often as those parts are sufficiently • agitated; whether that agitation be • made by Heat, or by Friction, or • Percussion, or Putrefaction, or by any vital Motion, • or any other Cause? As for instance; Sea-Water • in a raging Storm.î

7. Selected History of Bioluminescence, Continued • 1747 A.D Benjamin Franklin • First thought phosphorescence • was due to electricity. 1753 - "It is • indeed possible, that an extremely • small animalcule, too small to be visible even by the • best glasses, may yet give a visible light." • 1885 A.D. Raphael Dubois • 1952 A.D. E. Newton Harvey • ìThe Dean of Bioluminescenceî

8. Luciferase- The ìLight Bearingî Enzyme Luciferase is the general name applied to the class of enzymes that are responsible for converting chemical energy into light energy. The substrate molecule upon which luciferase acts is called a luciferin. While both luciferase and luciferin structures and their respective reactions vary widely between organisms, the requirement of atmospheric oxygen for bioluminescent activity remains universal for all of the various systems.

9. Light Cook Book (Mechanism of Bioluminescence) Luciferin Luciferase + + ATP + Mg2+ Luciferyladenylate-luciferase + Pyrophosphate Luciferyladenylate-luciferase + O2 Luciferase Oxyluciferin + + AMP + Light + CO2

10. Natural Supporting Cast of Bioluminescent Activity Organism group Member name Photo Vibrio harveyi Bacteria Polyporus Hanedai Fungi Dinoflagellates (Pyrocystis fusiformis) Protozoa

11. Natural Supporting Cast, Continued Organism group Member name Photo Phyllirrhoe (nudibranch) Mollusca Watasenia scintillans Cephalopoda Acholoe astericola Annelida No Picture Available Atolla vanhoeffeni Cnidaria

12. Natural Supporting Cast, Continued Organism group Member name Photo Arthropoda Euphausia pacifica (shrimp) Crustracea Pyrophorus noctilucus Insecta Flash Firefly Pediculati (Angler fish) Pisces

14. Applications of Bioluminescence • In Nature Photo compliments of Mike Hurley Obtained from http://www.coral.org/Gallery5.html • In the Lab

15. How and why some organisms use that funky bioluminescence in nature Reproduction • Pollination • Mating & Communication • Egg fertilization Predation • Lures • Mimicking Vision

16. Testing for TB Mycobacterium tuberculosis bacteria cause about 8 million cases of disease and million deaths a year. Time is a crucial factor for TB treatment. A new test making use of bioluminescence has been developed that can cut the time it takes to test for TB by about half.

17. The Procedure for the TB Test The luciferase (Lux) gene is inserted into a phage thatinfects the TB bacteria. The virus infects the cells and inserts the luciferase producing gene into the host genome. Luciferin substrate is added and the TB bacterium is now bioluminescent. The effect of different drugs on the TB is tested by adding the antibiotic to the transformed cells and detecting for light. If the antibiotic works, the light will not be emittedsince, of course, the cell is dead.

18. Bioluminescent X-Ray Specs Question: Wouldnít it be helpful (and neat!) to have a pair of glasses that would enable the wearer to see inside an animal in order to study diseases and infections? It may sound like something from a old comic book advertisement, but in reality, itís another new technique using bioluminescence to study organisms.

19. Infection Investigation the Cool Way Scientists have spliced a bacterial lux gene into salmonella. They then use this bacteria to infect mice and with little more than a Tim ìTheToolmanî Taylor enhanced camera, they can visualize and map the course of the infection.

20. Infection Investigation the Cool Way, Continued John Morrey created a transgenic mouse with the luciferase gene added to a genetic switch in every cell within its body The luciferase is turned on when DMSO, which simulates the effect of HIV in mice, is added to cause the switch to turn on. It is hoped that this animal can serve as the model for HIV infection or that this method can help gene therapy

21. Biosensors Biosensors are living organisms that are exploited for their ability to sense and report changes in their surroundings. The scientists that are working with these biosensors are using colonies of live bacteria that have a luciferase (Lux) gene tethered to another gene that is activated during an environmental change. When the change causes the native gene to be activated, the luciferase gene is also activated and light is produced, physically signaling the change.

22. Biosensors, Continued The key to the system lies in the special protein found on the surface of the bacterial cell wall. This special protein can be tailored to cause the bacteria to respond to different environmental cues. This is accomplished by mutating the surface protein producing gene. The mutants are then screened to identify the targets to which they are sensitive.

23. Toxin Luciferase Biosensor Mechanism SP Normal Gene Activated Lux gene activated

24. References Brown, Phyllida. Luciferin lights the way to faster TB tests. New Scientist.v. 138 May 15 '93 p. 16. Coghlan, Andy. Living sensors- Glowing bacteria can be tuned to detect almost anything. New Scientist.v. 158 no2134 May 16 '98 p. 16. Harvey, E. Newton. Bioluminescence. New York, 1952. Jacobs, William R., Jr; Barletta, Raul G; Udani, Rupa. Rapid assessment of drug susceptibilities of Mycobacterium tuberculosis by means of luciferase reporter phages. Science.v. 260 May 7 '93 p. 819-22. Meighen, Edward A. Molecular biology of bacterial bioluminescence. Microbiological Reviews.v. 55 Mar. '91 p. 123-42. Nelson, Wilfred H., Physical Methods for Microorganisms Detection. Boca Raton, 1991

25. References, Continued Siragusa GR, Nawotka K, Spilman SD, Contag PR, Contag CH. Real-time monitoring of escherichia coli O157:H7 adherence to beef carcass surface tissues with a bioluminescent reporter. Applied Environmental Microbiology 1999 Apr;65(4):1738-45. Taubes, Gary. Firefly gene lights up lab animals from inside out. Science.v. 276 June 27 '97 p. 1993. Travis, John. Following the inner light- Glow genes provide revealing pictures of infections. Science News.v. 150 Oct. 5 '96 p. 220-1. Bioluminescence Project by Ronney Abaza http://www.biology.lsa.umich.edu/~www/bio311/projects/ronney/biochem.shtml The Bioluminescence Web Page, Last updated: Mar 19, 1999 http://lifesci.ucsb.edu/~biolum/

27. Questions? Ask the Magic 8 BallÖ or meÖ itís your choice!