Revision as of 11:10, 2 August 2010

Luciferase

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.

[1] - Transformation protocol for students from basics - includes several plasmids

Plan
- set plasmid 555 into a BioBrick
- replace the promoter? sensitivity tuners?

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.

@@ Line 38: / Line 38: @@
 **set plasmid 555 into a BioBrick
 **replace the promoter? sensitivity tuners?
+*references
+** [http://www.pnas.org/content/84/19/6639.short Overproduction and purification of the luxR gene product: Transcriptional activator of the Vibrio fischeri luminescence system (Kaplan & Greenburg 1987) ]
+** [http://jb.asm.org/cgi/content/abstract/172/7/3974 Critical regions of the Vibrio fischeri luxR protein defined by mutational analysis (Slock et al 1990)]
 ==Transformation Experiment to test Firefly luciferase in registry==