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.
Bioluminescence Resonance Energy Transfer (BRET)
Bioluminescent molecules in gene expression
Identification of bacterial contamination in food
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.
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.
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.
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.
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.
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.
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.
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.
Methods of use
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
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.
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!
Artistic re-creation of the Bioluminescent Bay in Vieques
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
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.
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).
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)