Team:Imperial College London/Wetlab/BioBricks

From 2009.igem.org

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(Submitted BioBricks)
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| <partinfo>BBa_K200011 </partinfo>
| <partinfo>BBa_K200011 </partinfo>
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[[Image:II09_Composite.png|40px]]
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| 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.
| 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.

Revision as of 08:28, 14 October 2009



Submitted BioBricks

Imperial College iGEM 2009's uploaded BioBricks section on the Parts Registry can be found [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2009&group=Imperial%20College%20London here].

Registry Code Type Sequence Description
Coding

II09 Coding.png

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

II09 Coding.png

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

II09 Coding.png

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

II09 Coding.png

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

II09 Coding.png

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

II09 Coding.png

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

II09 Coding.png

[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

II09 Coding.png

[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

II09 Coding.png

[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

II09 Coding.png

[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

II09 Composite.png

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

II09 Composite.png

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.
Coding

II09 Coding.png

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

II09 Regulatory.png

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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