Difference between revisions of "Test pLsrA/cI"
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pBAD→RBS+lsrK+RBS+LsrR+TT | pBAD→RBS+lsrK+RBS+LsrR+TT | ||
| − | plsrA/ | + | plsrA/cI→RBS+GFP+TT does not glow |
[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_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] | ||
| Line 18: | Line 18: | ||
pBAD→RBS+lsrK+RBS+LsrR+TT | pBAD→RBS+lsrK+RBS+LsrR+TT | ||
| − | plsrA/ | + | plsrA/cI→RBS+GFP+TT 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_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] | ||
| Line 42: | Line 42: | ||
'''Ligation plan for test sequences''' | '''Ligation plan for test sequences''' | ||
| + | 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 pBAD with S/P, RBS+lsrK with X/P; do ligation. | ||
| + | |||
Digest TT with R/X, RBS+lsrR with R/S; 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 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. | Digest plsrA/cI with S/P, E0240 with X/P; do ligation. | ||
Latest revision as of 14:54, 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→RBS+GFP+TT does not glow
I13453+B0034+K091002+B0034+K091001+B0015
add AI-2
pBAD→RBS+lsrK+RBS+LsrR+TT
plsrA/cI→RBS+GFP+TT 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
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.
