Team:Cambridge/Voltage

From 2008.igem.org

(Difference between revisions)
(QvqFTCUcweZLILsFYdD)
 
(44 intermediate revisions not shown)
Line 1: Line 1:
-
<hr width="826px">
+
http://buy-ventolin.top/#8379 buy ventolin,
-
<div id="about">
+
-
{{Cambridge08a}}
+
-
</div>
+
-
<hr width="826px">
+
-
__NOTOC__
+
-
{|{{table}} width="820"
+
-
|-
+
-
|align="center" style="background-color: #444444;" |
+
-
[[Image:Voltage_button.gif‎ |300px]]
+
-
|colspan="1"|
+
-
 
+
-
{|cellspacing="5" cellpadding="10" style="background:#ffffff; width: 820px;"
+
-
|-valign="top"
+
-
|style="background:#ffffff"|
+
-
 
+
-
<font style="color:#000000">
+
-
 
+
-
=Background=
+
-
 
+
-
*The voltage output part of our project aims to mimic the signal transduction that occurs at a neural synapse.
+
-
*We are engineering E.coli to create a voltage output on detection of glutamate. This imitates the creation of a postsynaptic potential in a dendrite when a neurotransmitter (such as glutamate) is present at the synapse.
+
-
*The mechanism we have designed is similar to that used in the brain – relying on ion movement across the membrane, and gated ion channels.
+
-
*To simplify the concept, we are only regulating and measuring the flux of potassium (K+) ions, and we are using a directly glutamate-gated K+ ion channel.
+
-
*This means that on the binding of glutamate, the channels will open, allowing a K+ flux, which will change the voltage of the medium enough to be detected with a very sensitive electrode.
+
-
*In order to set up a large enough K+ concentration gradient across the membrane for ions to flow down when the channels open, cells are grown in high K+ medium (100mM) and resuspended in low K+ medium.
+
-
*However, E.coli also has a number of osmoregulatory systems which use relative K+ ion concentrations to control turgor. There are K+ leak channels (Kch and Kef) in the membrane, so we have chosen E.coli strains with mutations in these genes as our chassis.
+
-
 
+
-
 
+
-
=Experiment Summaries=
+
-
 
+
-
[[IGEM:Cambridge/2008/Notebook/Voltage/Output| Electrical Output]]
+
-
 
+
-
[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth|Mutant Growth Rates]]
+
-
 
+
-
[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations|Cytoplasmic K+ Concentrations]]
+
-
 
+
-
 
+
-
Parts Construction:
+
-
 
+
-
*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation|KDP]]
+
-
 
+
-
*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation|GluR0]]
+
-
 
+
-
*[[IGEM:Cambridge/2008/Extracted_Parts | Extracted Biobrick Parts]]
+
-
 
+
-
 
+
-
 
+
-
 
+
-
==Technical Information==
+
-
 
+
-
[[IGEM:Cambridge/2008/Notebook/Voltage/Gene Design|Gene Design]]
+
-
 
+
-
[[IGEM:Cambridge/2008/Notebook/Voltage/Flame Photometer Calibration|Flame Photometer Calibration]]
+
-
 
+
-
[[IGEM:Cambridge/2008/Notebook/Voltage/OD600 Calibration|OD600 (Cell Density) Calibration]]
+
-
 
+
-
[[IGEM:Cambridge/2008/Notebook/Voltage/Mutant Strains |Mutant Strains Information]]
+
-
 
+
-
 
+
-
=Useful Links=
+
-
 
+
-
[http://expasy.org/tools/ Protein prediction tools]
+
-
 
+
-
[http://www.uniprot.org/ Uniprot database]
+
-
 
+
-
[[Media:Voltage_project.ppt |Presentation]]
+
-
 
+
-
=Literature=
+
-
 
+
-
[http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1214631| Kdp operon diagram]
+
-
 
+
-
[http://www.jbc.org/cgi/content/abstract/276/13/9590|Kdp plasmid]
+
-
 
+
-
[http://www.springerlink.com/content/6042632827845551/ The Kdp-ATPase system and its regulation]
+
-
 
+
-
Potential Chassis: [http://cgsc.biology.yale.edu/Strain.php?ID=107402 |Strain JW1242-1]
+
-
[http://cgsc.biology.yale.edu/Strain.php?ID=107065 Strain JW0710-1]
+
-
 
+
-
[http://www.ncbi.nlm.nih.gov/pubmed/4942756 Kdp mutant - paper from 1971]
+
-
 
+
-
[http://www.shigen.nig.ac.jp/WGR/link/link_E.coli_e.html Worldwide E.coli Databases]
+
-
 
+
-
[http://jb.asm.org/cgi/content/abstract/188/5/1950 Characterisation of kdpD - 2005]
+
-
 
+
-
[http://jb.asm.org/cgi/content/full/180/19/5102 Investigations on Kdp Operon exp. & flux]
+
-
 
+
-
[http://dx.doi.org/10.1006/jmbi.2001.4884 Very interesting 2001 paper concerning Glutamate Channels]
+
-
 
+
-
[http://www.nature.com/nature/journal/v402/n6763/full/402817a0.html 1999 paper on functional characterization of prokaryote Glu Channels]
+
-
 
+
-
[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=BA000022 Sequenced Synechocystis PCC 6803 genome]
+
-
 
+
-
[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=47118304&from=1401809&to=1403002&view=gbwithparts  Glutamate-gated K+ channel GluR0]
+
-
 
+
-
[http://redpoll.pharmacy.ualberta.ca/CCDB/cgi-bin/STAT_NEW.cgi| Link to E.coli statistics page (CCDB Database)]
+
-
 
+
-
|-
+
-
|colspan="2" style="background-color: #F2F2F2;"|
+
-
{{LnNotebookRecentChanges2}}
+
-
|}
+
-
 
+
-
</font>
+
-
__NOEDITSECTION__
+
-
__NOTOC__
+

Latest revision as of 20:25, 4 May 2016

http://buy-ventolin.top/#8379 buy ventolin,

Retrieved from "http://2008.igem.org/Team:Cambridge/Voltage"