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Team:Imperial College London/Wetlab/BioBricks
From 2009.igem.org
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| - | | '''PAH+TT''' | + | | '''PAH+TT''' Ligation to a double terminator means that a closed transcriptional unit is produced once a promoter and RBS are ligated before this construct |
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| - | | '''pCstA+RBS+RcsB+TT''' | + | | '''pCstA+RBS+RcsB+TT''' This is the functional transcriptional unit which allows expression of the RcsB gene and production of colanic acid. |
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Revision as of 16:35, 18 October 2009
- Back to Hub
- Cloning Strategy
- Protocols
- BioBricks
- Results
- Notebook
Submitted BioBricks
Summary Table of BioBricks Designed
| Registry Code | Type | Sequence Description |
|---|---|---|
| Coding
| RcsB is a receiver protein which acts as a positive regulator of a number of genes including capsule genes responsible for colanic acid production. | |
| Coding
| Dam (DNA Adenine Methylase) The methylase encoded by the dam gene ([http://en.wikipedia.org/wiki/Dam_(methylase) Dam methylase]) transfers a methyl group from S-adenosylmethionine to the N6 position of the adenine residues in the sequence GATC, this protects the DNA from cleavage. | |
| Coding
| Colanic acid global regulator ygiV (B3023) increases the production of colanic acid further in conjunction with RcsB by acting as a repressor for mcbR/yncC promoter. YncC/mcbR normally repress colanic acid overproduction so as to increase biofilm formation. | |
| Coding
| Waal Ligase is an enzyme responsible for the ligation of an [http://en.wikipedia.org/wiki/O_antigen#O-antigen O-antigen] to the core [http://en.wikipedia.org/wiki/Oligosaccharide oligosaccharide] in the Gram-negative bacterium's outer membrane. | |
| Coding
| OtsA is the first of two required in the conversion of glucose to [http://en.wikipedia.org/wiki/Trehalose trehalose].
This enzyme catalyses the following reaction: UDP-glucose + D-glucose 6-phosphate -> UDP + alpha,alpha-trehalose 6-phosphate | |
| Coding
| OtsB This enzyme is the second of two required for the conversion of glucose to [http://en.wikipedia.org/wiki/Trehalose trehalose].
This enzyme catalyses the following reaction: alpha,alpha-trehalose 6-phosphate + H2O -> alpha,alpha-trehalose + phosphate | |
| Coding
| [http://en.wikipedia.org/wiki/Cellulase Cellulase] mainly catalyses the reactions that changes crystalline [http://en.wikipedia.org/wiki/Cellulose cellulose] to [http://en.wikipedia.org/wiki/Cellobiose cellobiose] and then finally to glucose. This cellulase is protease resistant. | |
| Coding
| [http://en.wikipedia.org/wiki/Phenylalanine_hydroxylase Phenylalanine hydroxylase] is the enzyme that breaks down [http://en.wikipedia.org/wiki/Phenylalanine phenylalanine] to [http://en.wikipedia.org/wiki/Tyrosine tyrosine]. Deficiency of this enzyme activity results in the autosomal recessive disorder [http://en.wikipedia.org/wiki/Phenylketonuria phenylketonuria]. | |
| Coding
| [http://en.wikipedia.org/wiki/Opiorphin Opiorphin] is a pentapeptide that inhibits the breakdown of endorphines. This results in powerful painrelief. This part contains an enterokinase cleavage site to facilitate synthesis and subsequent activation. | |
| Coding
| [http://en.wikipedia.org/wiki/Opiorphin Opiorphin] is a pentapeptide that inhibits the breakdown of endorphines. This results in powerful painrelief. This part contains an enterokinase cleavage site to facilitate synthesis and subsequent activation. This part also contains a HIS tag to facilitate high quality purification. | |
| Composite
| Restriction enzyme [http://www.thelabrat.com/restriction/DpnII.shtml DpnII] is a Type II restriction enzyme that recognises the sequence GATC. Its activity can be blocked by dam methylation. | |
| Composite
| Restriction enzyme [http://www.thelabrat.com/restriction/TaqI.shtml TaqI] is a Type II restriction enzyme that recognises the sequence TCGA. Its activity can be blocked by dam methylation. | |
| Composite
| This Lamda cI repressor has a cI857 mutation that results in denaturation of the repressor when the temperature is raised from 30 to 42°C, thereby allowing lambda promoter expression.
When the temperature is raised, typically to 42°C, the functionality of the protein is lost and the cI repressor is no longer able to bind to the operators on its promoter. Therefore, lambda promoter expression increases. | |
| Composite
| Lambda promoter (cIts responsive) is different from the common lambda promoter in that it is able to be repressed by the temperature sensitive cI protein (BBa_K200011). When it is not being repressed after 42°C induction, it acts as a strong promoter. | |
| Composite
| RBS+OtsB (Info here) | |
| Composite
| pCstA+RBS+GFP+TT (Info here) | |
| Composite
| pLacI+RBS+RFP+TT+pCstA+RBS+GFP+TT (Info here) | |
| Composite
| pCstA+RBS This is an intermediate construct that can be ligated to a coding region and terminator to produce a functional transcriptional unit. | |
| Composite
| pLacI+RBS This is an intermediate construct that can be ligated to a coding region and terminator to produce a functional transcriptional unit. | |
| Composite
| Heat-inducible system with GFP reporter Using the BioBrick part by Harvard '08. | |
| Composite
| PAH+TT Ligation to a double terminator means that a closed transcriptional unit is produced once a promoter and RBS are ligated before this construct | |
| Composite
| RcsB+TT Colanic acid producing gene (RcsB) attached to a double terminator. This is an intermediate construct assembled just prior to promoter ligation. | |
| Composite
| pCstA+RBS+RcsB+TT This is the functional transcriptional unit which allows expression of the RcsB gene and production of colanic acid. | |
| Composite
| RcsB+RBS+GFP+TT (Info here) | |
| Composite
| PAH+RBS+GFP+TT (Info here) | |
| Coding
| Protease resistant PAH This protease resistant PAH bears a mutation that changes serine 16 into glutamine. This has the effect of mimicking phosphorylation of the protein; modification which has been shown to be associated with protection against proteases ([http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1216923/ Døskeland et al., 1996]) | |
| Composite
| pLacI+RBS+RcsB+TT (Info here) | |
