Test pLsrA/cI
Test pLsrA/cI promoter
The pLsrA/cI promoter is constitutively off and is repressed by the lsrR protein. The addition of AI-2 should de-repress the lsrR operon causing it to glow. The addition of the cI protein should repress the promoter, causing it to not glow.
Test pLsrA/cI repression:
pBAD→RBS+lsrK+RBS+LsrR+TT
plsrA/cI→GFP does not glow
I13453+B0034+K091002+B0034+K091001+B0015
add AI-2
pBAD→RBS+lsrK+RBS+LsrR+TT
plsrA/cI→GFP glows
I13453+B0034+K091002+B0034+K091001+B0015
| LsrR | cI | AI-2 + GFP | AI-2 - GFP |
|---|---|---|---|
| 0 | 0 | 1 | 1 |
| 1 | 0 | 1 | 0 |
| 0 | 1 | 0 | 0 |
| 1 | 1 | 0 | 0 |
Ligation plan for test sequences
K091106 ligated with E0240
Digest RBS with S/P, lsrK with X/P; do ligation. Digest RBS with S/P, lsrR with X/P; do ligation. Digest pBAD with S/P, RBS+lsrK with X/P; do ligation. Digest TT with R/X, RBS+lsrR with R/S; do ligation. Digest RBS+lsrR+TT with R/X, pBAD+RBS+lsrK with R/S,; do ligation. Digest plsrA/cI with S/P, E0240 with X/P; do ligation.
