Difference between revisions of "Test XOR Hybrid Promoters"

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Test Tet repression:
+
[[Test Tet repression]]
  
 
Some promoter (pbad)→RBS+TetR+t+t+pMnt/Tet→RBS+GFP does not glows
 
Some promoter (pbad)→RBS+TetR+t+t+pMnt/Tet→RBS+GFP does not glows

Revision as of 18:55, 30 June 2008

This is a rough idea of how to test the four hybrid promoters for the XOR gate.


Test pLuxR/cI

luxR/CI hybrid promoter is constitutively off. Ligate with GFP on the 3’ end. If luxR is present first then add AI-1, the bacteria should glow green. Then add CI, the bacteria should not glow.

Some promoter→LuxR+t+t+pLuxR/cI→GFP does not glows


add AI-1

Some promoter→LuxR+t+t+pLuxR/cI→GFP glows

add AI-1

Some promoter→LuxR+cI+t+t+pLuxR/cI→GFP does not glow


LuxR cI AI-1 + GFP AI-1 - GFP
0 0 0 0
1 0 1 0
0 1 0 0
1 1 0 0



Test 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.

Some promoter→LacI+t+t+pLac/Mnt→GFP does not glow

Some promoter→Mnt+t+t+pLac/Mnt→GFP does not glow


add IPTG

Some promoter→LacI+t+t+pLac/Mnt→GFP glows


LacI Mnt IPTG + GFP IPTG - GFP
0 0 1 1
0 1 0 0
1 0 1 0
1 1 0 0



Test pLsrR/cI

pLsrR/cI is constitutively on and is repressed by LsrR. The addition of AI-2, which becomes phosphorylated by LsrK, represses LsrR causing the promoter to turn on. The addition of cI to should repress the promoter causing it to turn off.

Some promoter→LsrK+LsrR+t+t+pLsrR/cI→GFP does not glow


add AI-2

Some promoter→LsrK+LsrR+t+t+pLsrR/cI→GFP glows


add AI-2

Some promoter→LsrK+LsrR+cI+t+t+pLsrR/cI→GFP does not glows

LsrR cI AI-1 + GFP AI-1 - GFP
0 0 1 1
1 0 1 0
0 1 0 0
1 1 0 0



Test pMnt/Tet

The pMnt/TetR promoter is constitutively on and is repressed by the TetR protein. The addition of aTc should repress the TetR causing it to glow. The addition of the Mnt protein should repress the promoter causing it not glow.


TetR Mnt aTc + GFP aTc - GFP
0 0 1 1
0 1 0 0
1 0 1 0
1 1 0 0


Test Tet repression

Some promoter (pbad)→RBS+TetR+t+t+pMnt/Tet→RBS+GFP does not glows

I13453+RBS+C0040+TT+K091105+E0240


add aTc

Some promoter→TetR+t+t+pMnt/Tet→GFP glows

I13453+RBS+C0040+TT+K091105+E0240


Test Mnt repression:

Some promoter→Mnt+t+t+pMnt/Tet→GFP glows

I13453+RBS+C0072+TT+K091105+E0240


Test Tet and Mnt repression:

Some promoter→TetR+Mnt+t+t+pMnt/Tet→GFP does not glows

I13453+RBS+C0040+C0072+TT+K091105+E0240

add aTc does not glow