PMnt/Lac

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.