Team:PKU Beijing/Notebook/AND Gate 1/Input/LacI TetR 7

Some inducing data

Resource:
Delete LVA reverse tetR-tetP-GFP, colony, myself, renamed as DRTPG
Reverse tetR-tetP-GFP, colony, myself, RTPG
TetR-tetP-GFP, colony, myself (K228820), TPG
LacI-lacP-GFP, colony, myself (K228819), LPG

2009.8.17

Inducing:
Transfer five different colonies (DRTPG, RTPG, TPG, LPG, control) into 5 different tubes of 5mL LB.
When they grow to OD 0.4~0.6, we transfer 2mL LB of each tube into 2 ep tubes. Add some IPTG (1mM) or aTc (1mM) to one ep tube to induce, and the other one without any IPTG, as a control.
After 4 hours, spin down the Ecoli and pour out LB, and then suspend the Ecoli with PBS buffer.
Now we can put them under blue light to see the GFP.

Here is the result:

The upper row is control group (without inducing): from left to right is control, LPG, TPG, RTPG, DRTPG;
The under row is induced group: from left to right is control, LPG, TPG, RTPG, DRTPG;

Result & discussion

The result is consistent to the result of colonies on the plate (for more detail, refer to my prior experiment notes). The four devices LPG, TPG, RTPG, and DRTPG do not work very well, because the repressors (lacI & tetR) can not significantly repress the promoter lacP and tetP (or pLac and pTet).
We do not have too much time waste on them, so we decide to give up these four devices.

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