Team:Cambridge/References/ProjectBioluminescence

From 2010.igem.org

(Difference between revisions)
(Sequences)
 
(17 intermediate revisions not shown)
Line 1: Line 1:
-
{{:Team:Cambridge/Templates/header}}
+
{{:Team:Cambridge/Templates/headerMinimalprototype}}
-
{{:Team:Cambridge/LumNavTemplate}}
+
{{:Team:Cambridge/Templates/RefBar}}
-
=Bioluminescence=
+
{{:Team:Cambridge/Templates/headerbar|colour=#96d446|title=Bioluminescence: Introduction}}
 +
 
==The Plan==
==The Plan==
* Characterise the transfer function of PoPs + CHBT  --> Light
* Characterise the transfer function of PoPs + CHBT  --> Light
 +
* Our Characterisation should look like [http://www.nature.com/nbt/journal/v26/n7/abs/nbt1413.html this].
==The Theory==
==The Theory==
Line 9: Line 11:
[[Image:Cam-luci-cycle.jpg | 600px]]
[[Image:Cam-luci-cycle.jpg | 600px]]
-
==Increasing Light Emission==
+
==Amazing mutant==
-
*[http://www.ncbi.nlm.nih.gov/pubmed/12234677 Paper on isolation of G-LRE and H-LRE, which lists previous attempts to improve light production]
+
paper on cloning the lux operon from Ponyfish Photobacterium leiognathi into E.coli, by the E.coli mutant 43R the luminescence output could be increased dramatically to near native levels. --> find out what 43R does! [http://www3.interscience.wiley.com/cgi-bin/fulltext/120766166/PDFSTART Chan et al. 1991] suspects negative effectors in wild-type E.coli, but not in the mutant or in P.leiognathi, inhibit luminescence. If this is the case, it might be difficult to mimic the mutant phenotype with genes imported on a plasmid.  
-
*[http://www.ncbi.nlm.nih.gov/pubmed/10036167 Intensified light emission in presence of low ATP concentration by PPDK]
+
 
-
*[http://www.ncbi.nlm.nih.gov/pubmed/1332531 Enhanced luciferase activity through cytodine nucleotides]
+
[http://www.springerlink.com/content/vlxb2c59xut1htd8/fulltext.pdf Ulitzur et al. 1997] describes phenotypes like 43R being generated by deletions in the H-NS gene, fancy that. Apparently H-NS acts as a pleiotropic transcriptional repressor, with particularly strong effects on a certain set of promoters. Mutants have a reduced expression of a flagellar protein and are non-motile, but fully viable. In fact they appear to grow faster. However, H-NS has been implicated in E.coli thermostability.
-
*[http://www.ncbi.nlm.nih.gov/pubmed/12044905 Gene chimerisation to improve practical usefulness of firefly luciferase]
+
-
*[http://www.ncbi.nlm.nih.gov/pubmed/17540326 Genetically modified firefly luciferase - the EPIC luciferase]
+
-
==Sequences==
+
==ADLA==
-
*[http://www.ncbi.nlm.nih.gov/nucleotide/14331151?report=genbank&log$=nucltop&blast_rank=1&RID=4EUAXMJW01R A-LRE (P.pyralis) Sequence]
+
The Abrupt Decline in Luciferase Activity is a phenomenon described for bacterial luciferase in E.coli in stationary phase of growth (see [http://www.springerlink.com/content/w73k840k27866462/fulltext.pdf Koga et al. 2005]) The paper identifies two mutants, HupA and an N-terminal deletion of H-NS for which ADLA does not occur. hns205, the mutation necessary for quiescence is a C-terminal deletion. We should find out whether the phenotypes are the same, which would be sweet if we ended up doing quiescence as well.
-
*[http://www.ncbi.nlm.nih.gov/nuccore/AB261988.1 Mutant (EPIC) Luciferase (P.pyralis) Sequence]
+
<html>
 +
</div>
 +
</html>
 +
{{:Team:Cambridge/Templates/footerMinimal}}

Latest revision as of 12:16, 7 October 2010