Team:UIUC Illinois/Project/Design

Take up
The take up of L-carnitine is being amplified by the addition of Pseudomonas aeruginosa's L-carnitine uptake system. We desired to amplify this uptake system to allow for our bacteria to out compete our gutt bacteria. 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

After each gene was issolcated we ligated them 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 with an inducable T7 promoter.
"Break down" system: How to take up. Related plasmids design, explanation


When designing primers for this biobrick, we modified the gene. The gene originally began with a GTG sequence but was modified to ATG to improve the transcription factors recognition of the start codon. After gene isolation and digestion it was ligated into the pCDF which is another excellent expression vector and with an inducable T7 promoter.
Capsule? or other name?

How to take up. Related info