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

Kathryn Chung at 16:54, 8 September 2008

2008-09-08T16:54:31Z

←Older revision		Revision as of 16:54, 8 September 2008
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	=='''Transformation with episomal vectors'''==		=='''Transformation with episomal vectors'''==

-	'''ECE166''' : High-level constitutive expression of a Green Fluorescent Protein (with the promoter ~~PupP~~)	+	'''ECE166''' : High-level constitutive expression of a Green Fluorescent Protein (with the promoter Pupp)

	[[Image: phototrans166.jpg \| thumb \| 300px \| center \| Observation with microscope (x20) : B.S. transformed with ECE166 ]]		[[Image: phototrans166.jpg \| thumb \| 300px \| center \| Observation with microscope (x20) : B.S. transformed with ECE166 ]]

Yee Yen Tang at 16:31, 8 September 2008

2008-09-08T16:31:09Z

←Older revision		Revision as of 16:31, 8 September 2008
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	=Working with ''B.subtilis''=		=Working with ''B.subtilis''=
-	Building on the work of the last year's Cambridge iGEM team, we are exploring applications of the gram-positive chassis ''B.subtillis''. Easy to handle and transform, this bacterium is much better to use than ''E.coli'' wherever protein import/export is concerned, e.g. in our signalling project. An important part of our effort is to establish standard protocols and parts to work in ''B.subtillis'', characterise control elements, and develop new vectors. You can find our ~~''Bacillus'' protocols~~ [[IGEM:Cambridge/2008/Turing Pattern Formation/Experiments/Bacillus subtilis transfomation\|here]].	+	Building on the work of the last year's Cambridge iGEM team, we are exploring applications of the gram-positive chassis ''B.subtillis''. Easy to handle and transform, this bacterium is much better to use than ''E.coli'' wherever protein import/export is concerned, e.g. in our signalling project. An important part of our effort is to establish standard protocols and parts to work in ''B.subtillis'', characterise control elements, and develop new vectors. You can find our [[IGEM:Cambridge/2008/Turing Pattern Formation/Experiments/Bacillus subtilis transfomation\| ''Bacillus'' protocols here]].

	=Creating new Biobrick-compatible ''B.subtilis'' vectors=		=Creating new Biobrick-compatible ''B.subtilis'' vectors=

Yee Yen Tang at 15:12, 8 September 2008

2008-09-08T15:12:36Z

←Older revision		Revision as of 15:12, 8 September 2008
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	[[Image: 153Amylase.jpg \| thumb \| 200px \| left \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]		[[Image: 153Amylase.jpg \| thumb \| 200px \| left \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]
-	[[Image: 112Amylase.jpg \| thumb \| 200px \| ~~none~~ \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]	+	[[Image: 112Amylase.jpg \| thumb \| 200px \| center \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]

Yee Yen Tang at 15:12, 8 September 2008

2008-09-08T15:12:11Z

←Older revision		Revision as of 15:12, 8 September 2008
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	__NOEDITSECTION__		__NOEDITSECTION__
		+	__NOTOC__

Yee Yen Tang at 14:39, 8 September 2008

2008-09-08T14:39:19Z

←Older revision		Revision as of 14:39, 8 September 2008
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	[[Image: 153Amylase.jpg \| thumb \| 200px \| left \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]		[[Image: 153Amylase.jpg \| thumb \| 200px \| left \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]
	[[Image: 112Amylase.jpg \| thumb \| 200px \| none \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]		[[Image: 112Amylase.jpg \| thumb \| 200px \| none \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]
		+
		+

	=Testing ''B.subtilis'' promoters and RBSs=		=Testing ''B.subtilis'' promoters and RBSs=
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	# Transform ''B. subtilis'' with the plasmid which contains amyE for integration and lacZ for beta-galactosidase assay		# Transform ''B. subtilis'' with the plasmid which contains amyE for integration and lacZ for beta-galactosidase assay
	# Perform beta- galatosidase assay!		# Perform beta- galatosidase assay!
		+

	=Links=		=Links=
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	__NOEDITSECTION__		__NOEDITSECTION__
-	~~__NOTOC__~~

Yee Yen Tang at 14:38, 8 September 2008

2008-09-08T14:38:07Z

←Older revision		Revision as of 14:38, 8 September 2008
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	\|-		\|-
	\|}		\|}
		+
		+

	=Successful transformation of Bacillus=		=Successful transformation of Bacillus=

Chapart Marie at 16:27, 6 September 2008

2008-09-06T16:27:17Z

←Older revision		Revision as of 16:27, 6 September 2008
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	'''ECE153''' and '''ECE112''' : Transformation in IA751 : Amylase test		'''ECE153''' and '''ECE112''' : Transformation in IA751 : Amylase test

-	[[Image: 153Amylase.jpg \| thumb \| 200px \| ~~center~~ \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]	+	[[Image: 153Amylase.jpg \| thumb \| 200px \| left \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]
-	[[Image: 112Amylase.jpg \| thumb \| 200px \| ~~center~~ \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]	+	[[Image: 112Amylase.jpg \| thumb \| 200px \| none \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]

	=Testing ''B.subtilis'' promoters and RBSs=		=Testing ''B.subtilis'' promoters and RBSs=

Chapart Marie at 16:23, 6 September 2008

2008-09-06T16:23:23Z

←Older revision		Revision as of 16:23, 6 September 2008
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	'''ECE153''' and '''ECE112''' : Transformation in IA751 : Amylase test		'''ECE153''' and '''ECE112''' : Transformation in IA751 : Amylase test

-	~~{\| class="wikitable" border="0" align="center"~~	+	[[Image: 153Amylase.jpg \| thumb \| 200px \| center \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]
-	[[Image: 153Amylase.jpg \| thumb \| 200px \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]	+	[[Image: 112Amylase.jpg \| thumb \| 200px \| center \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]
-	[[Image: 112Amylase.jpg \| thumb \| 200px \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]	+
-	\|-	+
-	\|}	+

	=Testing ''B.subtilis'' promoters and RBSs=		=Testing ''B.subtilis'' promoters and RBSs=

Chapart Marie at 16:21, 6 September 2008

2008-09-06T16:21:00Z

←Older revision		Revision as of 16:21, 6 September 2008
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	=Successful transformation of Bacillus=		=Successful transformation of Bacillus=
		+	=='''Transformation with episomal vectors'''==
		+
		+	'''ECE166''' : High-level constitutive expression of a Green Fluorescent Protein (with the promoter PupP)
		+
		+	[[Image: phototrans166.jpg \| thumb \| 300px \| center \| Observation with microscope (x20) : B.S. transformed with ECE166 ]]
		+
		+	=='''Transformation with integration vectors'''==
		+
		+	'''ECE153''' and '''ECE112''' : Transformation in IA751 : Amylase test
		+
		+	{\| class="wikitable" border="0" align="center"
		+	[[Image: 153Amylase.jpg \| thumb \| 200px \| IA751 transformed with ECE153 : SUCCESFUL TRANSFORMATION, no zone of clearing ]]
		+	[[Image: 112Amylase.jpg \| thumb \| 200px \| IA751 transformed with ECE112 : SUCCESFUL TRANSFORMATION for 4 colonies out of 5, no zone of clearing ]]
		+	\|-
		+	\|}

	=Testing ''B.subtilis'' promoters and RBSs=		=Testing ''B.subtilis'' promoters and RBSs=

Chapart Marie at 16:17, 6 September 2008

2008-09-06T16:17:14Z

←Older revision		Revision as of 16:17, 6 September 2008
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	\|-		\|-
	\|}		\|}
		+
		+	=Successful transformation of Bacillus=
		+
	=Testing ''B.subtilis'' promoters and RBSs=		=Testing ''B.subtilis'' promoters and RBSs=
	We are seeking to test the expression strength and response to inducers in several promoters in ''B.subtilis'', in combination with different Bacillus-specific ribosome binding sites.		We are seeking to test the expression strength and response to inducers in several promoters in ''B.subtilis'', in combination with different Bacillus-specific ribosome binding sites.