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Team:UIUC Illinois/Project/Design
From 2013.igem.org
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| - | <br><b>"Take up" and "Break down"</b> | + | <br><b>"Take up"</b><br> |
| + | The take up of L-carnitine is being amplified by the addition of Pseudomonas aeruginosa's L-carnitine uptake system. This system consisted of two genes cbcVW and caiX. cbcVW is the membrane bound protein that allows for L-carnitine to go through the bacteria's membrane. caiX is a free protein that directs the L-carnitine to the membrane bound transporter. We have introduced both of these genes in the Nissle and tested the uptake of L-carnitine. | ||
| + | <b>" Break down" </b> | ||
| + | The break down is achieved by cdhCAB also originating from the Pseudomonas aeruginosa's genome. This enzyme breaks down L-carnitine into 3-dehydrocarnitine. This process allows, as we mentioned on the Background page, for the alternative breakdown of L-carnitine diverting TMAO production. | ||
| + | <br> | ||
<br><b> "Take up" system:</b> How to take up. Related plasmids design, explanation<br><br> | <br><b> "Take up" system:</b> How to take up. Related plasmids design, explanation<br><br> | ||
| - | + | They were placed into two different plasmids. cbcVW was added into pET 26b and caiX was added into pACYC. These vectors were selected because of its excellent expression capabilities and an inducable T7 promoter. | |
<br><b> "Break down" system:</b> How to take up. Related plasmids design, explanation<br><br><br> | <br><b> "Break down" system:</b> How to take up. Related plasmids design, explanation<br><br><br> | ||
| - | + | This gene originally began with a GTG sequence but this was modified to ATG to improve the transcription factors recognition of the start codon. After PCR was ligated into the pCDF which is another excellent expression vector and an inducable T7 promoter. | |
<br><b> Capsule? or other name?</b><br><br> | <br><b> Capsule? or other name?</b><br><br> | ||
Revision as of 02:56, 28 September 2013
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"Take up"
The take up of L-carnitine is being amplified by the addition of Pseudomonas aeruginosa's L-carnitine uptake system. This system consisted of two genes cbcVW and caiX. cbcVW is the membrane bound protein that allows for L-carnitine to go through the bacteria's membrane. caiX is a free protein that directs the L-carnitine to the membrane bound transporter. We have introduced both of these genes in the Nissle and tested the uptake of L-carnitine.
" Break down"
The break down is achieved by cdhCAB also originating from the Pseudomonas aeruginosa's genome. This enzyme breaks down L-carnitine into 3-dehydrocarnitine. This process allows, as we mentioned on the Background page, for the alternative breakdown of L-carnitine diverting TMAO production.
"Take up" system: How to take up. Related plasmids design, explanation
They were placed into two different plasmids. cbcVW was added into pET 26b and caiX was added into pACYC. These vectors were selected because of its excellent expression capabilities and an inducable T7 promoter.
"Break down" system: How to take up. Related plasmids design, explanation
This gene originally began with a GTG sequence but this was modified to ATG to improve the transcription factors recognition of the start codon. After PCR was ligated into the pCDF which is another excellent expression vector and an inducable T7 promoter.
Capsule? or other name?
How to take up. Related info
