Difference between revisions of "Test pMnt/Lac"
| Line 15: | Line 15: | ||
Some promoter→RBS+LacI+Mnt+t+t+pLac/Mnt→GFP does not glow | Some promoter→RBS+LacI+Mnt+t+t+pLac/Mnt→GFP does not glow | ||
| − | http://partsregistry.org/Part:BBa_I13453 I13453] → [http://partsregistry.org/Part:BBa_B0034 B0034] +LacI+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+Mnt+TT+ [http://partsregistry.org/Part:BBa_K091104 K091104]→[http://partsregistry.org/Part:BBa_E0240 E0240] |
'''To Test the promoter and addition of IPTG''' | '''To Test the promoter and addition of IPTG''' | ||
Revision as of 16:38, 8 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+Mnt+t+t+pLac/Mnt→GFP does not glow
I13453 → B0034 +LacI+Mnt+TT+ K091104→E0240
To Test the promoter and addition of IPTG
add IPTG
Some promoter→RBS+IPTG+LacI+t+t+pLac/Mnt→GFP glows
I13453 → B0034 +IPTG+LacI+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
