Difference between revisions of "Test pMnt/Lac"
| Line 3: | Line 3: | ||
'''To Test repression of promoter''' | '''To Test repression of promoter''' | ||
| − | Some promoter→RBS+LacI+t+t+pLac/Mnt→RBS+GFP | + | Some promoter→RBS+LacI+t+t+pLac/Mnt→RBS+GFP <nowiki>''does not glow''</nowiki> |
[http://partsregistry.org/Part:BBa_I13453 I13453] → [http://partsregistry.org/Part:BBa_B0034 B0034] +LacI+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | [http://partsregistry.org/Part:BBa_I13453 I13453] → [http://partsregistry.org/Part:BBa_B0034 B0034] +LacI+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | ||
| − | Some promoter→RBS+Mnt+t+t+pLac/Mnt→GFP | + | Some promoter→RBS+Mnt+t+t+pLac/Mnt→GFP <nowiki>''does not glow''</nowiki> |
[http://partsregistry.org/Part:BBa_I13453 I13453] → [http://partsregistry.org/Part:BBa_B0034 B0034] +Mnt+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | [http://partsregistry.org/Part:BBa_I13453 I13453] → [http://partsregistry.org/Part:BBa_B0034 B0034] +Mnt+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | ||
| − | Some promoter→RBS+LacI+RBS+Mnt+t+t+pLac/Mnt→GFP | + | Some promoter→RBS+LacI+RBS+Mnt+t+t+pLac/Mnt→GFP <nowiki>''does not glow''</nowiki> |
[http://partsregistry.org/Part:BBa_I13453 I13453] → [http://partsregistry.org/Part:BBa_B0034 B0034] +LacI+RBS+Mnt+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | [http://partsregistry.org/Part:BBa_I13453 I13453] → [http://partsregistry.org/Part:BBa_B0034 B0034] +LacI+RBS+Mnt+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | ||
| Line 21: | Line 21: | ||
add IPTG | add IPTG | ||
| − | Some promoter→IPTG+RBS+LacI+t+t+pLac/Mnt→GFP | + | Some promoter→IPTG+RBS+LacI+t+t+pLac/Mnt→GFP <nowiki>''glows''</nowiki> |
[http://partsregistry.org/Part:BBa_I13453 I13453] → IPTG+[http://partsregistry.org/Part:BBa_B0034 B0034]+LacI+TT+[http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | [http://partsregistry.org/Part:BBa_I13453 I13453] → IPTG+[http://partsregistry.org/Part:BBa_B0034 B0034]+LacI+TT+[http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | ||
| Line 27: | Line 27: | ||
| − | Some promoter→IPTG+RBS+LacI+Mnt+t+t+pLac/Mnt→GFP | + | Some promoter→IPTG+RBS+LacI+Mnt+t+t+pLac/Mnt→GFP <nowiki>'' glows''</nowiki> |
[http://partsregistry.org/Part:BBa_I13453 I13453] →IPTG+ [http://partsregistry.org/Part:BBa_B0034 B0034] +LacI+RBS+Mnt+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | [http://partsregistry.org/Part:BBa_I13453 I13453] →IPTG+ [http://partsregistry.org/Part:BBa_B0034 B0034] +LacI+RBS+Mnt+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] | ||
Latest revision as of 16:58, 16 July 2008
pMnt/Lac is constitutively on but is repressed by the LacI protein. The promoter is also repressed by the Mnt protein. The addition of IPTG should repress the LacI repressor and glow in the absence of the Mnt protein.
To Test repression of promoter
Some promoter→RBS+LacI+t+t+pLac/Mnt→RBS+GFP ''does not glow''
I13453 → B0034 +LacI+TT+ K091104→E0240
Some promoter→RBS+Mnt+t+t+pLac/Mnt→GFP ''does not glow''
I13453 → B0034 +Mnt+TT+ K091104→E0240
Some promoter→RBS+LacI+RBS+Mnt+t+t+pLac/Mnt→GFP ''does not glow''
I13453 → B0034 +LacI+RBS+Mnt+TT+ K091104→E0240
To Test the promoter and addition of IPTG
add IPTG
Some promoter→IPTG+RBS+LacI+t+t+pLac/Mnt→GFP ''glows''
I13453 → IPTG+B0034+LacI+TT+K091104→E0240
Some promoter→IPTG+RBS+LacI+Mnt+t+t+pLac/Mnt→GFP '' glows''
I13453 →IPTG+ B0034 +LacI+RBS+Mnt+TT+ K091104→E0240
| LacI | Mnt | IPTG + GFP | IPTG - GFP |
|---|---|---|---|
| 0 | 0 | 1 | 1 |
| 0 | 1 | 0 | 0 |
| 1 | 0 | 1 | 0 |
| 1 | 1 | 0 | 0 |
Steps to Build Hybrid Promoter
6/30/08
Perform a Ligation of Mnt/Lac (K091104) with E0240 (RBS + GFP)
Transform ligation over night
7/1/08
Clones were picked and set over night in LB + AMP
