Team:Cambridge/Bioluminescence/Bacterial Luciferases

From 2010.igem.org

(Difference between revisions)
 
(8 intermediate revisions not shown)
Line 1: Line 1:
{{:Team:Cambridge/Templates/headerMinimalprototype}}
{{:Team:Cambridge/Templates/headerMinimalprototype}}
{{:Team:Cambridge/Templates/headerbar|colour=#386abc|title=Project Vibrio: Introduction}}
{{:Team:Cambridge/Templates/headerbar|colour=#386abc|title=Project Vibrio: Introduction}}
-
 
-
 
'''Project Vibrio''' was designed to create BioBricks from genes involved in bacterial bioluminescence.  
'''Project Vibrio''' was designed to create BioBricks from genes involved in bacterial bioluminescence.  
Line 13: Line 11:
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.
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 the 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].  
+
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==
==Our work==
{{:Team:Cambridge/Templates/RightImage|image=Phosphoreum_bright.jpg|caption=Our workspace illuminated by'' Vibrio phosphoreum'', a bacterium we investigated}}
{{:Team:Cambridge/Templates/RightImage|image=Phosphoreum_bright.jpg|caption=Our workspace illuminated by'' Vibrio phosphoreum'', a bacterium we investigated}}
-
To complement 'Project Firefly', we wanted to use lux operons for the following two purposes:
+
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.
* 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.
* 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].
 
{{:Team:Cambridge/Templates/footer}}
{{:Team:Cambridge/Templates/footer}}

Latest revision as of 03:00, 28 October 2010