IGEM:Cambridge/2008/Notebook/Voltage - Revision history

Ellis O'Neill at 13:38, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 13:38:07 GMT

←Older revision		Revision as of 13:38, 28 October 2008
Line 40:		Line 40:

	[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]		[[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:		Parts Construction:

-	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]	+	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

-	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]	+	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

-	[[IGEM:Cambridge/2008/~~Notebook/Voltage/K+ Growth~~\|~~Mutant Growth Rates~~]]	+	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]

-	~~[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]~~

-	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]

	=Progress=		=Progress=

Ellis O'Neill at 13:37, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 13:37:19 GMT

←Older revision		Revision as of 13:37, 28 October 2008
Line 43:		Line 43:
	Parts Construction:		Parts Construction:

-	**[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]	+	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

-	**[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]	+	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]
Line 51:		Line 51:
	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]

-	**[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]	+	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]

	=Progress=		=Progress=

Ellis O'Neill at 13:36, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 13:36:54 GMT

←Older revision		Revision as of 13:36, 28 October 2008
Line 41:		Line 41:
	[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]		[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]

-	Parts Construction	+	Parts Construction:

-	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]	+	**[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

-	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]	+	**[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]
Line 51:		Line 51:
	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]

-	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]	+	**[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]

	=Progress=		=Progress=

Ellis O'Neill at 13:36, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 13:36:20 GMT

←Older revision		Revision as of 13:36, 28 October 2008
Line 41:		Line 41:
	[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]		[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]

-	~~[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]~~	+	Parts Construction

-	[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]	+	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]
		+
		+	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]

Ellis O'Neill at 11:14, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 11:14:26 GMT

←Older revision		Revision as of 11:14, 28 October 2008
Line 39:		Line 39:
	=Experiment Summaries=		=Experiment Summaries=

-	[IGEM:Cambridge/2008/Notebook/Voltage/Output\|Electrical Output]	+	[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]

	[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]		[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

Ellis O'Neill at 11:14, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 11:14:05 GMT

←Older revision		Revision as of 11:14, 28 October 2008
Line 38:		Line 38:

	=Experiment Summaries=		=Experiment Summaries=
		+
		+	[IGEM:Cambridge/2008/Notebook/Voltage/Output\|Electrical Output]

	[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]		[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

Ellis O'Neill at 11:13, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 11:13:12 GMT

←Older revision		Revision as of 11:13, 28 October 2008
Line 36:		Line 36:
	*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.		*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.

-	~~[[Media:Voltage_project.ppt \|Presentation]]~~

-	=~~Progress~~=	+	=Experiment Summaries=

-	[[IGEM:Cambridge/2008/Notebook/Voltage/~~Progress~~ \|~~Progress~~]]	+	[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

-	~~=Experiment Summaries=~~	+	[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]

	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]
-
-	~~[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]~~

	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]		*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]

-	[[IGEM:Cambridge/2008/Notebook/Voltage/~~GluR0 Manipulation~~\|~~GluR0~~]]	+	=Progress=
		+
		+	[[IGEM:Cambridge/2008/Notebook/Voltage/Progress \|Progress]]


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	[http://www.uniprot.org/ Uniprot database]		[http://www.uniprot.org/ Uniprot database]
		+
		+	[[Media:Voltage_project.ppt \|Presentation]]

	=Literature=		=Literature=

Ellis O'Neill at 10:53, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 10:53:19 GMT

←Older revision		Revision as of 10:53, 28 October 2008
Line 48:		Line 48:
	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]

-	[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|~~BioBrick Manipulation~~]]	+	[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]		*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]
		+
		+	[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

Ellis O'Neill at 10:26, 28 October 2008

Ellis O'Neill — Tue, 28 Oct 2008 10:26:36 GMT

←Older revision		Revision as of 10:26, 28 October 2008
Line 32:		Line 32:
	*The mechanism we have designed is similar to that used in the brain – relying on ion movement across the membrane, and gated ion channels.		*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.		*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.
-	~~[[Image:Voltage_schematic.png \|thumb \|left\| 350px \| Concept schematic]]~~
	*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.		*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, ~~an ion pump is necessary. E.coli has a transmembrane P-type ATPase called Kdp, which pumps~~ K+ ~~into the cell. We have isolated this gene to overexpress it, therefore causing the cells to pump~~ in ~~a large number of~~ K+ ~~ions~~.	+	*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 ~~ordered~~ E.coli strains with mutations in these genes ~~to allow K+ to remain sequestered inside the cells until the glutamate-gated channels open~~.	+	*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.

	[[Media:Voltage_project.ppt \|Presentation]]		[[Media:Voltage_project.ppt \|Presentation]]

Rebecca Koenigsberg-Miles at 11:27, 12 September 2008

Rebecca Koenigsberg-Miles — Fri, 12 Sep 2008 11:27:41 GMT

←Older revision		Revision as of 11:27, 12 September 2008
Line 32:		Line 32:
	*The mechanism we have designed is similar to that used in the brain – relying on ion movement across the membrane, and gated ion channels.		*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.		*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.
-	[[Image:Voltage_schematic.png \|thumb \|left\| ~~300px~~ \| Concept schematic]]	+	[[Image:Voltage_schematic.png \|thumb \|left\| 350px \| Concept schematic]]
	*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.		*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, an ion pump is necessary. E.coli has a transmembrane P-type ATPase called Kdp, which pumps K+ into the cell. We have isolated this gene to overexpress it, therefore causing the cells to pump in a large number of K+ ions.		*In order to set up a large enough K+ concentration gradient across the membrane for ions to flow down when the channels open, an ion pump is necessary. E.coli has a transmembrane P-type ATPase called Kdp, which pumps K+ into the cell. We have isolated this gene to overexpress it, therefore causing the cells to pump in a large number of K+ ions.

←Older revision		Revision as of 13:38, 28 October 2008
Line 40:		Line 40:

	[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]		[[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:		Parts Construction:

-	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]	+	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

-	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]	+	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

-	[[IGEM:Cambridge/2008/~~Notebook/Voltage/K+ Growth~~\|~~Mutant Growth Rates~~]]	+	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]

-	~~[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]~~

-	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]

	=Progress=		=Progress=

←Older revision		Revision as of 13:36, 28 October 2008
Line 41:		Line 41:
	[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]		[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]

-	Parts Construction	+	Parts Construction:

-	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]	+	**[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

-	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]	+	**[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]
Line 51:		Line 51:
	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]

-	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]	+	**[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]

	=Progress=		=Progress=

←Older revision		Revision as of 13:36, 28 October 2008
Line 41:		Line 41:
	[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]		[[IGEM:Cambridge/2008/Notebook/Voltage/Output\| Electrical Output]]

-	~~[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]~~	+	Parts Construction

-	[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]	+	*[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]
		+
		+	*[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]

	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Growth\|Mutant Growth Rates]]

←Older revision		Revision as of 10:53, 28 October 2008
Line 48:		Line 48:
	[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]		[[IGEM:Cambridge/2008/Notebook/Voltage/K+ Concentrations\|Cytoplasmic K+ Concentrations]]

-	[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|~~BioBrick Manipulation~~]]	+	[[IGEM:Cambridge/2008/Notebook/Voltage/BioBrick Manipulation\|KDP]]

	*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]		*[[IGEM:Cambridge/2008/Extracted_Parts \| Extracted Biobrick Parts]]
		+
		+	[[IGEM:Cambridge/2008/Notebook/Voltage/GluR0 Manipulation\|GluR0]]