"
Team:Cambridge/References/ProjectBioluminescence/Luciferase
From 2010.igem.org
(Difference between revisions)
(→Transformation Experiment to test Firefly luciferase in registry) |
|||
| Line 2: | Line 2: | ||
{{:Team:Cambridge/LumNavTemplate}} | {{:Team:Cambridge/LumNavTemplate}} | ||
| - | + | =Luciferase= | |
==Mutant luciferase (P. pyralis)== | ==Mutant luciferase (P. pyralis)== | ||
| Line 10: | Line 10: | ||
==Other Insect luciferases== | ==Other Insect luciferases== | ||
| - | * For colours we could use the click beetle luciferase [http://partsregistry.org/wiki/index.php/Part:BBa_J70005 red] or [http://partsregistry.org/wiki/index.php/Part:BBa_J70006 green]. They're reliable and from the Knight lab but not in the distribution. | + | *For colours we could use the click beetle luciferase [http://partsregistry.org/wiki/index.php/Part:BBa_J70005 red] or [http://partsregistry.org/wiki/index.php/Part:BBa_J70006 green]. They're reliable and from the Knight lab but not in the distribution. |
==Bacterial Bioluminescence== | ==Bacterial Bioluminescence== | ||
Revision as of 14:34, 26 July 2010
Luciferase
Mutant luciferase (P. pyralis)
- Mutant Enhanced Photon Initiating Complex (EPIC) Luciferase (P.pyralis) Sequence
- Site-directed mutagenesis of firefly luciferase active site amino acids: a proposed model for bioluminescence color
- Improved practical usefulness of firefly luciferase by gene chimerization and random mutagenesis
Other Insect luciferases
- For colours we could use the click beetle luciferase red or green. They're reliable and from the Knight lab but not in the distribution.
Bacterial Bioluminescence
- Vibrio Fischeri
- Vibrio Harveyi
- Genetics of luxCDABE (adapted from wikipedia)
- 2006 attempts at BioBricking the lux operon
- Notes
- The lux wiki from parts registry
- NCBI Lux operon sequence
- parts registry lux operon -currently waiting on an email to see how useful/available this part is
- IMPORTANT INFO
- The Photorhabdus luminescens luxCDABE operon in the registry. It's not well characterised
Plasmid experiment
- Vibrio plasmid experiment -possible source of lux operon
- [1] - Transformation protocol for students from basics - includes several plasmids
- Plan
- set plasmid 555 into a BioBrick
- replace the promoter? sensitivity tuners?
Transformation Experiment to test Firefly luciferase in registry
- Firefly luciferase is in the registry as BBa_I712019, and have been sent to us as part of the spring 2010 distribution.
- We need to find a new vector with a bacterial ribosyme binding site, initiator, and a stop codon. Having searched via the catalog in the plasmids section, then in expression plasmids, we have selected a constitutive plasmid:
- The plasmid selected is BBa_J13002, which is a TetR repressed POPS/RIPS generator, again part of the spring 2010 distribution - kit plate 1, well 13B, plasmid pSB1A2. It was proven to work by UMN iGEM 2009.
