Difference between revisions of "Test Mnt/Tet promoter"
| Line 4: | Line 4: | ||
| − | Test tet repression | + | '''1) 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 | ||
| Line 17: | Line 17: | ||
| − | Test Mnt repression: | + | '''2) Test Mnt repression:''' |
Some promoter→Mnt+t+t+pMnt/Tet→GFP glows | Some promoter→Mnt+t+t+pMnt/Tet→GFP glows | ||
| Line 24: | Line 24: | ||
| − | Test Tet and Mnt repression: | + | '''3) Test Tet and Mnt repression:''' |
Some promoter→TetR+Mnt+t+t+pMnt/Tet→GFP does not glows | Some promoter→TetR+Mnt+t+t+pMnt/Tet→GFP does not glows | ||
| Line 34: | Line 34: | ||
'''06/30/08''' | '''06/30/08''' | ||
| + | |||
K091105 ligated with E0240 | K091105 ligated with E0240 | ||
| − | need to ligate some promoter with an RBS and ligate TetR with a double terminator | + | 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 those two parts together | ||
then ligate that part with the Mnt/Tet with GFP | then ligate that part with the Mnt/Tet with GFP | ||
Revision as of 19:26, 30 June 2008
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
