Difference between revisions of "Test pLsrA/cI"
| Line 6: | Line 6: | ||
''' Test pLsrA/cI repression:''' | ''' Test pLsrA/cI repression:''' | ||
| − | pBAD→RBS+LsrR+TT | + | pBAD→RBS+lsrK+RBS+LsrR+TT |
plsrA/cI→GFP does not glow | plsrA/cI→GFP does not glow | ||
| − | [http://partsregistry.org/Part:BBa_I13453 I13453]+[http://partsregistry.org/Part:BBa_B0034 B0034]+[http://partsregistry.org/Part:BBa_K091001 K091001]+[http://partsregistry.org/Part:BBa_B0015 B0015] | + | [http://partsregistry.org/Part:BBa_I13453 I13453]+[http://partsregistry.org/Part:BBa_B0034 B0034]+[http://partsregistry.org/Part:BBa_K091002 K091002]+[http://partsregistry.org/Part:BBa_B0034 B0034]+[http://partsregistry.org/Part:BBa_K091001 K091001]+[http://partsregistry.org/Part:BBa_B0015 B0015] |
| + | |||
| + | [http://partsregistry.org/Part:BBa_K091106 K091106]+[http://partsregistry.org/Part:BBa_E0240 E0240] | ||
add AI-2 | add AI-2 | ||
| − | pBAD→RBS+LsrR+TT | + | pBAD→RBS+lsrK+RBS+LsrR+TT |
| + | |||
| + | plsrA/cI→GFP glows | ||
| − | + | [http://partsregistry.org/Part:BBa_I13453 I13453]+[http://partsregistry.org/Part:BBa_B0034 B0034]+[http://partsregistry.org/Part:BBa_K091002 K091002]+[http://partsregistry.org/Part:BBa_B0034 B0034]+[http://partsregistry.org/Part:BBa_K091001 K091001]+[http://partsregistry.org/Part:BBa_B0015 B0015] | |
| − | + | [http://partsregistry.org/Part:BBa_K091106 K091106]+[http://partsregistry.org/Part:BBa_E0240 E0240] | |
{| border="1" cellpadding="2" | {| border="1" cellpadding="2" | ||
| Line 41: | Line 45: | ||
K091106 ligated with E0240 | 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. | ||
Revision as of 14:51, 11 July 2008
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.
