Imperial College/Courses/Spring2008/Synthetic Biology/Team 5 - Revision history

Tom Adie at 16:21, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 16:21:59 GMT

←Older revision		Revision as of 16:21, 11 March 2008
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	Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.		Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.

-	There is already a lacZ reporter in the [http://parts.mit.edu MIT registry]; part [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0033 BBa_E0033]. This part is a section of the lacZ gene (the lacZ-alpha fragment) such that, when cultured with N-terminal deletion mutants of lacZ, enzyme activity is restored and plates can be assayed by eye. This is standard for the lacZ reporter construct as it saves a large number of valuable base pairs. As it currently stands, however, this part is [http://parts.mit.edu/registry/index.php/Part:BBa_E0033:Experience missing an in-~~line~~ stop codon] (as verified by sequencing of the physical part) and so may be non-functional.	+	There is already a lacZ reporter in the [http://parts.mit.edu MIT registry]; part [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0033 BBa_E0033]. This part is a section of the lacZ gene (the lacZ-alpha fragment) such that, when cultured with N-terminal deletion mutants of lacZ, enzyme activity is restored and plates can be assayed by eye. This is standard for the lacZ reporter construct as it saves a large number of valuable base pairs. As it currently stands, however, this part is [http://parts.mit.edu/registry/index.php/Part:BBa_E0033:Experience missing an in-frame stop codon] (as verified by sequencing of the physical part) and so may be non-functional.

	== Our system so far: ==		== Our system so far: ==

	[[Image:System Overview.jpg\|600px\|Overview of our system]]		[[Image:System Overview.jpg\|600px\|Overview of our system]]

Tom Adie at 16:20, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 16:20:22 GMT

←Older revision		Revision as of 16:20, 11 March 2008
Line 24:		Line 24:
	Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.		Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.

-	There is already a lacZ reporter in the [http://parts.mit.edu MIT registry]; part [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0033 BBa_E0033]. This part is a section of the lacZ gene (the lacZ-alpha fragment) such that, when cultured with N-terminal deletion mutants of lacZ, enzyme activity is restored and plates can be assayed by eye.	+	There is already a lacZ reporter in the [http://parts.mit.edu MIT registry]; part [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0033 BBa_E0033]. This part is a section of the lacZ gene (the lacZ-alpha fragment) such that, when cultured with N-terminal deletion mutants of lacZ, enzyme activity is restored and plates can be assayed by eye. This is standard for the lacZ reporter construct as it saves a large number of valuable base pairs. As it currently stands, however, this part is [http://parts.mit.edu/registry/index.php/Part:BBa_E0033:Experience missing an in-line stop codon] (as verified by sequencing of the physical part) and so may be non-functional.

	== Our system so far: ==		== Our system so far: ==

	[[Image:System Overview.jpg\|600px\|Overview of our system]]		[[Image:System Overview.jpg\|600px\|Overview of our system]]

Tom Adie at 16:01, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 16:01:10 GMT

←Older revision		Revision as of 16:01, 11 March 2008
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	Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.		Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.
		+
		+	There is already a lacZ reporter in the [http://parts.mit.edu MIT registry]; part [http://parts.mit.edu/registry/index.php?title=Part:BBa_E0033 BBa_E0033]. This part is a section of the lacZ gene (the lacZ-alpha fragment) such that, when cultured with N-terminal deletion mutants of lacZ, enzyme activity is restored and plates can be assayed by eye.

	== Our system so far: ==		== Our system so far: ==

	[[Image:System Overview.jpg\|600px\|Overview of our system]]		[[Image:System Overview.jpg\|600px\|Overview of our system]]

Tom Adie at 15:19, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 15:19:17 GMT

←Older revision		Revision as of 15:19, 11 March 2008
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	== Our system so far: ==		== Our system so far: ==

-	[[Image:System Overview.jpg\|~~500px~~\|Overview of our system]]	+	[[Image:System Overview.jpg\|600px\|Overview of our system]]

Tom Adie at 15:16, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 15:16:45 GMT

←Older revision		Revision as of 15:16, 11 March 2008
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	== Our system so far: ==		== Our system so far: ==

-	[[Image:System Overview.jpg\|~~200px~~\|Overview of our system]]	+	[[Image:System Overview.jpg\|500px\|Overview of our system]]

Tom Adie at 15:16, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 15:16:26 GMT

←Older revision		Revision as of 15:16, 11 March 2008
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	== Our system so far: ==		== Our system so far: ==

-	[[Image:System Overview.jpg]]	+	[[Image:System Overview.jpg\|200px\|Overview of our system]]

Tom Adie at 15:15, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 15:15:58 GMT

←Older revision		Revision as of 15:15, 11 March 2008
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	== Our system so far: ==		== Our system so far: ==

-	[[Image:~~Timpski1~~]]	+	[[Image:System Overview.jpg]]

Tom Adie at 15:09, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 15:09:53 GMT

←Older revision		Revision as of 15:09, 11 March 2008
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	== Our system so far: ==		== Our system so far: ==

-	[[Image:~~System Overview.jpg~~]]	+	[[Image:Timpski1]]

Tom Adie at 15:07, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 15:07:33 GMT

←Older revision		Revision as of 15:07, 11 March 2008
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	Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.		Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.
		+
		+	== Our system so far: ==
		+
		+	[[Image:System Overview.jpg]]

Tom Adie at 14:42, 11 March 2008

Tom Adie — Tue, 11 Mar 2008 14:42:21 GMT

←Older revision		Revision as of 14:42, 11 March 2008
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	While in testing, however, we need an easily-detectable reporter. GFP as a candidate was swiftly dismissed, as using a fluorescent protein to report on the activity of light-sensitive bacteria would undoubtedly lead to interference and skewed results! Antibiotic expression in concert with antibiotic resistance could be relevant as antibiotic-producing bacteria may be a useful bio-implant and would also be an accurate reporter - the complexity of this system may impose on our timetable, however, so we have decided ultimately to use the well-characterised lacZ reporter construct. In this system, bacteria that produce the protein exhibit a blue colouration as opposed to a standard white colour for negative results.		While in testing, however, we need an easily-detectable reporter. GFP as a candidate was swiftly dismissed, as using a fluorescent protein to report on the activity of light-sensitive bacteria would undoubtedly lead to interference and skewed results! Antibiotic expression in concert with antibiotic resistance could be relevant as antibiotic-producing bacteria may be a useful bio-implant and would also be an accurate reporter - the complexity of this system may impose on our timetable, however, so we have decided ultimately to use the well-characterised lacZ reporter construct. In this system, bacteria that produce the protein exhibit a blue colouration as opposed to a standard white colour for negative results.

-	http://upload.wikimedia.org/wikipedia/commons/4/48/Lac_operon1.png [http://en.wikipedia.org/wiki/Image:Lac_operon1.png]<br>	+	http://upload.wikimedia.org/wikipedia/commons/4/48/Lac_operon1.png [http://en.wikipedia.org/wiki/Image:Lac_operon1.png]<br><br>

	Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.		Thus if we tack on a lacZ reporter to the construct the system could be tested using plated cells exposed to light patterns and examined later for distribution of blue colonies vs. white ones.