Difference between revisions of "PMnt/Lac"
From GcatWiki
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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. | 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. | ||
| − | Plan to Test the promoter | + | '''Plan to Test the promoter''' |
| − | To Test repression using LacI | + | '''To Test repression using LacI''' |
Some promoter→LacI+t+t+pLac/Mnt→GFP does not glow | Some promoter→LacI+t+t+pLac/Mnt→GFP does not glow | ||
| − | To Test repression using Mnt protein | + | '''To Test repression using Mnt protein''' |
Some promoter→Mnt+t+t+pLac/Mnt→GFP does not glow | Some promoter→Mnt+t+t+pLac/Mnt→GFP does not glow | ||
| − | To Test pLac being on with addition of IPTG | + | '''To Test pLac being on with addition of IPTG''' |
add IPTG | add IPTG | ||
| Line 21: | Line 21: | ||
| − | To Test the repression of pLac | + | '''To Test the repression of pLac''' |
Some promoter→LacI+Mnt+t+t+pLac/Mnt→GFP does not glow | Some promoter→LacI+Mnt+t+t+pLac/Mnt→GFP does not glow | ||
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| − | 6-30-08 Ligation | + | '''6-30-08 Ligation''' |
Mnt/ Lac [http://partsregistry.org/Part:BBa_K091104 K091104] was ligated to [http://partsregistry.org/Part:BBa_E0240 E0240] | Mnt/ Lac [http://partsregistry.org/Part:BBa_K091104 K091104] was ligated to [http://partsregistry.org/Part:BBa_E0240 E0240] | ||
The plasmid + vector was then transformed onto an amp plate. | The plasmid + vector was then transformed onto an amp plate. | ||
Latest revision as of 20:09, 30 June 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.
Plan to Test the promoter
To Test repression using LacI
Some promoter→LacI+t+t+pLac/Mnt→GFP does not glow
To Test repression using Mnt protein
Some promoter→Mnt+t+t+pLac/Mnt→GFP does not glow
To Test pLac being on with addition of IPTG
add IPTG
Some promoter→LacI+t+t+pLac/Mnt→GFP glows
To Test the repression of pLac
Some promoter→LacI+Mnt+t+t+pLac/Mnt→GFP does not glow
| LacI | Mnt | IPTG + GFP | IPTG - GFP |
|---|---|---|---|
| 0 | 0 | 1 | 1 |
| 0 | 1 | 0 | 0 |
| 1 | 0 | 1 | 0 |
| 1 | 1 | 0 | 0 |
6-30-08 Ligation
Mnt/ Lac K091104 was ligated to E0240
The plasmid + vector was then transformed onto an amp plate.
