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Team:Imperial College London/Biobricks
From 2009.igem.org
[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2009&group=Imperial%20College%20London Biobricks]
| Registry Code | Type | Sequence Description |
|---|---|---|
| Coding | RcsB is a receiver protein in a two component phosphorelay system, which acts as a positive regulator of a number of genes including capsule genes responsible for colanic acid production. This is via the activation of the ugd/cps operon which is required for capsule synthesis. | |
| 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. When methylation occurs in the recognition site of a particular group of restriction endonuclease including MboI, 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 O-antigen to the core oligosaccharide in the Gram-negative bacterium's outer membrane.
Unlike other exopolysaccharides, colanic acid does not naturally bind to the cell surface but rather forms a thick mesh between cells. While Waal Ligase usually links the O-antigen to the core oligosaccharide, in K-12 it links colanic acid to the core oligosaccharide. | |
| Coding | OtsA is the first of two required in the conversion of glucose to 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 trehalose.
This enzyme catalyses the following reaction: alpha,alpha-trehalose 6-phosphate + H2O -> alpha,alpha-trehalose + phosphate | |
| Coding | Cellulase mainly catalyses the reactions that changes crystalline cellulose to cellobiose and then finally to glucose. It also catalyses, to a small extent, the break down of carboxymethyl cellulose. This cellulase is protease resistant. | |
| Coding | 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 | Restriction enzyme DpnII is a Type II restriction enzyme that recognises the sequence GATC. Its activity can be blocked by dam methylation.
The working temperature is 37°C. | |
| Coding | Restriction enzyme TaqI is a Type II restriction enzyme that recognises the sequence TCGA. Its activity can be blocked by dam methylation.
| |
| Coding | 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.
The repressor normally negatively regulates the expression of genes from the bacteriophage lambda pL and pR promoters. This repressive action is strongest at 30°C. However, 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. | |
| Regulatory | 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
It has been shown through lysis experiments that this lambda promoter can be stringently repressed by the cI protein at temperatures up to 30°C. At temperatures higher than 30°C, the cI repressor is progressively thermally denatured and no longer able to bind to the promoter operator regions. Therefore, promoter activity is induced. | |
