Transcript No Slide Title

Dave Quiles Imagineering Productions Presents
An Illuminating
Presentation
In Illumicolor
(Where Available)
Flash
Firefly
Davey
Q
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.
Selected History of Bioluminescence
ï500 B.C. Aniximenes
He described light exhibited by the sea when struck
with an oar.
ï350 B.C. Aristotle
Considered lightning similar to when "you strike the
sea with a rod by night and the water is seen to
shine".
ï1637 A.D. Rene Descartes
Striking seawater will "generate
sparks rather similar to those which
are emitted by pieces of flint when
they are struck."
Selected History of Bioluminescence, Continued
ï1668 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
ï1717 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
Selected History of Bioluminescence, Continued
ï1747 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."
ï1885 A.D. Raphael Dubois
ï1952 A.D. E. Newton Harvey
ìThe Dean of Bioluminescenceî
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.
Light Cook Book (Mechanism of Bioluminescence)
Luciferin + Luciferase + ATP + Mg2+
Luciferyladenylate-luciferase + Pyrophosphate
Luciferyladenylate-luciferase + O2
Luciferase + Oxyluciferin + AMP + Light + CO2
Natural Supporting Cast of Bioluminescent
Activity
Organism group
Member name
Photo
Bacteria
Vibrio harveyi
Fungi
Polyporus Hanedai
Protozoa
Dinoflagellates
(Pyrocystis fusiformis)
Natural Supporting Cast, Continued
Organism group
Member name
Photo
Mollusca
Phyllirrhoe
(nudibranch)
Cephalopoda
Watasenia
scintillans
Annelida
Acholoe
astericola
Cnidaria
Atolla
vanhoeffeni
No Picture
Available
Natural Supporting Cast, Continued
Organism group
Member name
Arthropoda
Crustracea
Euphausia pacifica
(shrimp)
Insecta
Pyrophorus
Flash Firefly
noctilucus
Pisces
Pediculati
(Angler fish)
Photo
Applications of Bioluminescence
ïIn Nature
Photo compliments of Mike Hurley
Obtained from http://www.coral.org/Gallery5.html
ïIn the Lab
How and why some organisms use that funky
bioluminescence in nature
Reproduction
ïPollination
ïMating & Communication
ïEgg fertilization
Predation
ïLures
ïMimicking
Vision
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.
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.
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.
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.
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
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.
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.
Biosensor Mechanism
Toxin
SP
Normal
Gene
Activated
Luciferase
Lux gene
activated
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
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.sht
ml
The Bioluminescence Web Page, Last updated: Mar 19, 1999
http://lifesci.ucsb.edu/~biolum/
Questions?
Ask the Magic 8 BallÖ
or meÖ
itís your choice!