Difference between revisions of "Test pLsrA/cI"
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− | ''' | + | ''' Test pLsrA/cI repression:''' |
− | + | pBAD→RBS+LsrR+TT | |
− | + | plsrA/cI→GFP does not glow | |
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[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_K091106 K091106]+[http://partsregistry.org/Part:BBa_E0240 E0240] | [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_K091106 K091106]+[http://partsregistry.org/Part:BBa_E0240 E0240] | ||
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add AI-2 | add AI-2 | ||
− | pBAD→RBS+LsrR+TT | + | pBAD→RBS+LsrR+TT |
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+ | plsrA/cI→GFP glows | ||
[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_K091106 K091106]+[http://partsregistry.org/Part:BBa_E0240 E0240] | [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_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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− | '''Ligation plan for | + | '''Ligation plan for test sequences''' |
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K091106 ligated with E0240 | K091106 ligated with E0240 | ||
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then ligate that part 5' to the plsrA/cI+GFP | then ligate that part 5' to the plsrA/cI+GFP | ||
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Revision as of 16:01, 10 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+LsrR+TT
plsrA/cI→GFP does not glow
I13453+B0034+K091001+B0015+K091106+E0240
add AI-2
pBAD→RBS+LsrR+TT
plsrA/cI→GFP glows
I13453+B0034+K091001+B0015+K091106+E0240
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
need to ligate some promoter (digest with E/S) with an RBS (digest E/X) and ligate lsrR(digest with E/S) with a double terminator (digest with E/X)
then ligate the promoter+RBS to the 5' end of the lsrR+TT parts together
then ligate that part 5' to the plsrA/cI+GFP