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.
1) Test LsrR repression:
pBAD→RBS+LsrR+t+t+plsrA/cI→RBS+GFP does not glow
I13453+B0034+K091001+B0015+K091106+E0240
add AI-2
pBAD→RBS+TetR+TT+plsrA/cI→GFP glows
I13453+B0034+K091001+B0015+K091106+E0240
2) Test cI repression:
pBAD→RBS+cI+TT+plsrA/cI→GFP glows
I13453+B0034+C0051+B0015+K091106+E0240
add cI protein
pBAD→RBS+cI+TT+plsrA/cI→GFP does not glow
I13453+B0034+C0051+B0015+K091106+E0240
3) Test LsrR and cI repression:
pBAD→RBS+lsrR+cI+TT+plsrA/cI→GFP does not glows
I13453+B0034+K091001+C0051+B0015+K091106+E0240
add AI-2
pBAD→RBS+lsrR+cI+TT+plsrA/cI→GFP does not glows
I13453+B0034+K091001+C0051+B0015+K091106+E0240
LsrR | cI | AI-2 + GFP | AI-2 - GFP |
---|---|---|---|
0 | 0 | 1 | 1 |
0 | 1 | 0 | 0 |
1 | 0 | 1 | 0 |
1 | 1 | 0 | 0 |