Test Mnt/Tet promoter

Test Mnt/Tet promoter

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.

1) 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

2) Test Mnt repression:

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

I13453+RBS+C0072+TT+K091105+E0240

3) 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

06/30/08

K091105 ligated with E0240

need to ligate some promoter (digest with E and S) with an RBS (digest E and X) and ligate TetR(digest with E and S) with a double terminator (digest with E and x)

then ligate those two parts together

then ligate that part with the Mnt/Tet with GFP