Difference between revisions of "Test pLsrA/cI"
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| − | ''' | + | ''' Test pLsrA/cI repression:''' |
| − | pBAD→RBS+ | + | pBAD→RBS+lsrK+RBS+LsrR+TT |
| − | + | 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_K091106 K091106]+[http://partsregistry.org/Part:BBa_E0240 E0240] | |
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| + | add AI-2 | ||
| − | + | pBAD→RBS+lsrK+RBS+LsrR+TT | |
| − | + | plsrA/cI→RBS+GFP+TT glows | |
| − | I13453+B0034+ | + | [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] | |
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{| border="1" cellpadding="2" | {| border="1" cellpadding="2" | ||
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|- | |- | ||
| 0 || 0 || 1 || 1 | | 0 || 0 || 1 || 1 | ||
| + | |- | ||
| + | | 1 || 0 || 1 || 0 | ||
|- | |- | ||
| 0 || 1 || 0 || 0 | | 0 || 1 || 0 || 0 | ||
| − | |||
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|- | |- | ||
| 1 || 1 || 0 || 0 | | 1 || 1 || 0 || 0 | ||
|} | |} | ||
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| + | '''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. | ||
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.
