Test XOR Hybrid Promoters
This is a rough idea of how to test the four hybrid promoters for the XOR gate.
Test pMnt/Lac
pMnt/Lac is constitutively on but is repressed by the LacI protein. The promoter is also repressed by the Mnt protein. The addition of IPTG should repress the LacI repressor and glow in the absence of the Mnt protein.
Some promoter→LacI+t+t+pLac/Mnt→GFP does not glow
Some promoter→Mnt+t+t+pLac/Mnt→GFP does not glow
add IPTG
Some promoter→LacI+t+t+pLac/Mnt→GFP glows
| LacI | Mnt | IPTG + GFP | IPTG - GFP |
|---|---|---|---|
| 0 | 0 | 1 | 1 |
| 0 | 1 | 0 | 0 |
| 1 | 0 | 1 | 0 |
| 1 | 1 | 0 | 0 |
The pMnt/TetR promoter is constitutively on and is repressed by the TetR protein. The addition of aTc should repress the TetR causing it to glow. The addition of the Mnt protein should repress the promoter causing it not glow.
| TetR | Mnt | aTc + GFP | aTc - GFP |
|---|---|---|---|
| 0 | 0 | 1 | 1 |
| 0 | 1 | 0 | 0 |
| 1 | 0 | 1 | 0 |
| 1 | 1 | 0 | 0 |
Test tet repression
Some promoter (pbad)→RBS+TetR+t+t+pMnt/Tet→RBS+GFP does not glows
I13453+RBS+C0040+TT+K091105+E0240
add aTc
Some promoter→TetR+t+t+pMnt/Tet→GFP glows
I13453+RBS+C0040+TT+K091105+E0240
Test Mnt repression:
Some promoter→Mnt+t+t+pMnt/Tet→GFP glows
I13453+RBS+C0072+TT+K091105+E0240
Test Tet and Mnt repression:
Some promoter→TetR+Mnt+t+t+pMnt/Tet→GFP does not glows
I13453+RBS+C0040+C0072+TT+K091105+E0240
add aTc does not glow
