Team:Imperial College London/Biobricks

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[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.
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.

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.

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