Team:Exeter/Results/biobricks

From 2012.igem.org

Revision as of 16:54, 26 September 2012

ExiGEM

Concept

Beyond
The
Science

Results

• Introduction
• Team
• Collaborators

• Background & Concept
• Future Applications
• Human Practices

• Project Conclusions
• Safety
• Diary
• Lab Books
• Bio-Bricks
• Achievements

• Early Ideas
• Outreach

Follow our Blog
Follow us on Twitter

<BIOBRICKS SUBMITTED TO THE REGISTRY

Due to the time available we were only able to get 14 of our potential Biobricks into pSB1C3 and sent into the registry. Over 30 more were waiting for there chance. Our project was certainly worth more than the sum of it's Registry parts!</font>

Encodes the galactosytransferase wbiP that adds a β-D-Galactose donor sugar onto an α-D-GalNAc(N-Acetylgalactosamine) acceptor sugar to form a 1->3 glycosidic bond. wbiP readily glycosylates GalNAc analogues all with α substutions at the reducing end, and also glycosylates serine and threonine residues. It's close relatives include: WbnJ from Escherichia coli O86, wcmC from Escherichia coli O86, wbgO from Escherichia coli 055:H7, wfdH from Escherichia coli 0167, wbsK from Escherichia coli O128 and wfbZ from Escherichia coli O147.

Sequence and Features

<partinfo>BBa_K764007</partinfo>

Encodes the glucoronic acid transferase wfcA that adds a β-D-GlcA donor sugar onto an α-D-Mannose acceptor sugar to form a 1->2 glycosidic bond. It's close relatives include: wfeE from Escherichia coli O32, WfaO from Escherichia coli O24, wbhT from Escherichia coli O22 and wbaZ from Escherichia coli O9:K30.

Sequence and Features

<partinfo>BBa_K764003</partinfo>

Encodes the fucosyltransferase wbnK that adds a branched α-L-Fucose donor sugar onto a β-D-Galactose acceptor sugar to form a 1->2 glycosidic bond. The protein wbnK has strict selectivity for Gal-β-(1->3)-GalNAc and acceptor specificity assays have shown that a change of glycosidic bond from 1->3 to 1->4 or from the β to the α configuration both lead to poor substrates for wbnK. wbnK is involved in the formation of both the O86 O-antigen and human blood group B antigen tetrasaccharide. It's close relatives include: wbwK from Escherichia coli O86:B7, wbgL from Escherichia coli O126, wbgN from Escherichia coli O55:H7 and wbgN from Escherichia coli O55:H6.

Sequence and Features

<partinfo>BBa_K764002</partinfo>

Encodes the galactosytransferase wbnJ that adds a β-D-Galactose donor sugar onto an α-D-GalNAc(N-Acetylgalactosamine) acceptor sugar to form a 1->3 glycosidic bond. wbnJ is thought to be involved in colanic acid biosynthesis, acting as an acetyltransferase. It's close relatives include: wcmC from Escherichia coli O86:K2:H2, wfdH from Escherichia coli O167, wbsK from Escherichia coli O128, wbnA from Escherichia coli O113 and wbwJ from Escherichia coli O86:B7.

Sequence and Features

<partinfo>BBa_K764000</partinfo>

Encodes the glycosyltransferase wclY from Escherichia coli O107 that adds an α-D-GlcNAc (N-Acetylglucosamine) donor sugar onto a β-D-Galactose acceptor sugar to form a 1->4 glycosidic bond. wclY has been identified as the glycosyltransferase that gives the differences between Escherichia coli O107 and O117 O-antigens. It's close relatives include: wclY from Escherichia coli O117 and wfeK from Escherichia coli O112ac.

Sequence and Features

<partinfo>BBa_K764004</partinfo>

The complete construct of Newcastle iGEM 2010's biobrick BBa_K322921 for SacB. This was linked to the signal peptide for export from the cell and prevent damage to our E coli.

Sequence and Features

<partinfo>BBa_K764033</partinfo>

Sequence and Features

<partinfo>BBa_K764034</partinfo>

Sequence and Features

<partinfo>BBa_K764026</partinfo>

The enzyme hyaluronan synthase is responsible for generating hyaluronan from D-glucuronic acid and D-N-acetylglucosamine, linked via alternating β-1,4 and β-1,3 glycosidic bonds.

Sequence and Features

<partinfo>BBa_K764025</partinfo>

<partinfo>BBa_K764024</partinfo>

Encodes hyaluronan synthase (hasA) that performs the following two reactions:

UDP-α-N-acetyl-D-glucosamine + β-D-glucuronosyl-(1->3)-N-acetyl-β-D-glucosaminyl-(1->4)-(nascent hyaluronan) = UDP + N-acetyl-β-D-glucosaminyl-(1->4)-β-D-glucuronosyl-(1->3)-N-acetyl-β-D-glucosaminyl-(1->4)-(nascent hyaluronan)

UDP-α-D-glucuronate + N-acetyl-β-D-glucosaminyl-(1->4)-β-D-glucuronosyl-(1->3)-(nascent hyaluronan) = UDP + β-D-glucuronosyl-(1->3)-N-acetyl-β-D-glucosaminyl-(1->4)-β-D-glucuronosyl-(1->3)-(nascent hyaluronan).

This generates hyaluronan, a virulence factor in streptococcus pyogenes if linked to lipid A molecules in the cell wall. Hyaluronan is also present in human joints and skin as a lubricant.

Sequence and Features

<partinfo>BBa_K764024</partinfo>

Performs the enzymatic conversion of starch into variable sized cyclic oligosaccharides. This glycosyltransferase (cgt) from Bacillis sp 1011 produces alpha-cyclodextrin, a six membered ring of α-D-glucopyranoside units linked 1->4 glycosidic bonds.

Sequence and Features

<partinfo>BBa_K764023</partinfo>

A construct of wbnK (BBa_K764002) attached to a (double) terminator (BBa_B0014) using 3A assembly. The function of wbnK remains unperturbed.

Sequence and Features

<partinfo>BBa_K764009</partinfo>

A construct of wclY (BBa_K764004) attached to a (double) terminator (BBa_B0014) using 3A assembly. The function of wclY remains unperturbed.

Sequence and Features

<partinfo>BBa_K764011</partinfo>