Team:Cambridge/Bioluminescence/Bacterial Luciferases

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Latest revision as of 03:00, 28 October 2010

Project Vibrio: Introduction

Project Vibrio was designed to create BioBricks from genes involved in bacterial bioluminescence.

Natural bioluminescent bacteria

Wave glowing with bioluminescent micro-organisms

Bioluminescence is a trait found in a number of marine bacteria. Some strains use their ability to emit light to form symbiotic relationships. A number of deep sea fish and squids have specialised light organs which harbour populations of bacteria that help their hosts by emitting light. One such example is the partnership between the Hawaiian Bobtail squid (Euprymna scolopes) and the bacterium Vibrio fischeri.

At night squid hunt high in the water column, attacking their prey from above. The squid uses the light produced by its symbionts to hide the shadow it casts when hunting in top waters in clear moonlit nights. An elaborate light sensing and shutter system adjusts the light output to the light that falls on the squid's back.

Bioluminescent bacterial species can differ markedly in their lifestyles. Vibrio Harveyi is a free living marine bacterium, while Xenorhabdus luminescens is a symbiont of terrestrial nematodes.

The light-generating chemical reactions in bioluminescent bacteria are catalysed by enzymes expressed from so-called lux genes. These genes encode not only the bacterial luciferase, but also the enzymes required for the synthesis of fatty aldehydes, which are the substrates for the reaction. In the bacterial genome, the lux genes occur clustered in the lux operon.

Our work

Our workspace illuminated by Vibrio phosphoreum, a bacterium we investigated

To complement 'Project Firefly', we intended to use the lux operon from Vibrio fischeri for the following three purposes:

Emission of blue light to complete our spectrum of emission wavelengths.
Substrate production within E. coli, avoiding the need for addition of external substrates, such as luciferin.
Design of a biosensor output device that can be combined with various input systems.

Bacterial lux operons encode five enzymes involved in the light-generating pathway. In nature, the lux genes appear to be repressed by the nucleoid protein, H-NS, and occur under quorum sensing control. We wished to relieve repression by H-NS to achieve brighter light outputs. We furthermore removed quorum sensing control to facilitate use of the part in biosensors under different regulatory inputs.

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 {{:Team:Cambridge/Templates/headerbar|colour=#386abc|title=Project Vibrio: Introduction}}
-{{:Team:Cambridge/Templates/LeftImage|image=Cambridge-Photobacterium_plate.JPG|caption=An agar plate with bioluminescent bacteria.}}
+'''Project Vibrio''' was designed to create BioBricks from genes involved in bacterial bioluminescence.
-'''Project Vibrio''' was designed to create BioBricks from prokaryotic genes encoding enzymes involved in the catalysis of light-generating chemical reactions. These genes can be found in the lux operons of bioluminescent bacteria.
 ==Natural bioluminescent bacteria==
-Bioluminescence is a trait found in a number of marine bacteria.  Some strains use their ability to emit light to form symbiotic relationships.  A number of deep sea fish and squids have specialised <em>light organs</em> which harbour populations of bacteria that help their hosts by emitting light.  One such example is the partnership between the Hawaiian Bobtail squid <em>(Euprymna scolopes)</em> and the bacterium <em>Vibrio fischeri</em>. At night squid hunt high in the water column, attacking their prey from above.  Vibrio Fischeri is usually found in symbiosis with the Hawaiian Bobtail Squid. The squid uses the light produced by its symbionts to hide the shadow it casts when hunting in top waters in clear moonlit nights. An elaborate light sensing and shutter system adjusts the light output to the light that falls on the squid's back. Other bioluminescent bacterial species can differ markedly in their lifestyles. Vibrio Harveyi is a free living marine bacterium, while Xenorhabdus luminescens is a symbiont of terrestrial nematodes.
+{{:Team:Cambridge/Templates/RightImage|image=Tide.jpg|caption=Wave glowing with bioluminescent micro-organisms}}
+Bioluminescence is a trait found in a number of marine bacteria.  Some strains use their ability to emit light to form symbiotic relationships.  A number of deep sea fish and squids have specialised <em>light organs</em> which harbour populations of bacteria that help their hosts by emitting light.  One such example is the partnership between the Hawaiian Bobtail squid <em>(Euprymna scolopes)</em> and the bacterium <em>Vibrio fischeri</em>.
+At night squid hunt high in the water column, attacking their prey from above.   The squid uses the light produced by its symbionts to hide the shadow it casts when hunting in top waters in clear moonlit nights. An elaborate light sensing and shutter system adjusts the light output to the light that falls on the squid's back.
+Bioluminescent bacterial species can differ markedly in their lifestyles. Vibrio Harveyi is a free living marine bacterium, while Xenorhabdus luminescens is a symbiont of terrestrial nematodes.
+The light-generating chemical reactions in bioluminescent bacteria are catalysed by enzymes expressed from so-called lux genes. These genes encode not only the bacterial luciferase, but also the enzymes required for the synthesis of fatty aldehydes, which are the substrates for the reaction. In the bacterial genome, the lux genes occur clustered in the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background lux operon].
 ==Our work==
-To complement 'Project Firefly', we wanted to use lux operons for the following two purposes:
+{{:Team:Cambridge/Templates/RightImage|image=Phosphoreum_bright.jpg|caption=Our workspace illuminated by'' Vibrio phosphoreum'', a bacterium we investigated}}
+To complement 'Project Firefly', we intended to use the lux operon from ''Vibrio fischeri'' for the following three purposes:
 * Emission of blue light to complete our spectrum of emission wavelengths.
 * Substrate production within E. coli, avoiding the need for addition of external substrates, such as luciferin.
+* Design of a biosensor output device that can be combined with various input systems.
+Bacterial lux operons encode five enzymes involved in the light-generating pathway. In nature, the lux genes appear to be repressed by the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background nucleoid protein, H-NS], and occur under [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background quorum sensing control]. We wished to relieve repression by H-NS to achieve brighter light outputs. We furthermore removed quorum sensing control to facilitate use of the part in biosensors under different regulatory inputs.
-Bacterial lux operons encode five enzymes involved in the light-generating pathway.
-*<i>luxA</i> and <i>luxB</i> form the luciferase of the system, which causes the emission of light when acting on the substrate tetradecanal.
-*<i>luxC</i>, <i>luxD</i> and <i>luxE</i> are involved in the biosynthesis of tetradecanal from readily available substrates.
-In nature, the lux genes appear to be repressed by the [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background nucleoid protein, H-NS], and occur under [https://2010.igem.org/Team:Cambridge/Bioluminescence/Background quorum sensing control].
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