From 2007.igem.org

Contents 1 Purpose 1: 2 Purpose 2: 3 Samples: 4 Procedure:

Purpose 1:

To check if LacI hybrid promoter is activated by AHL and repressed by LacI.

Purpose 2:

To obtain parameters of LacI hybrid promoter for computational simulation.

Samples:

1. pTrc99A + [LacI hybrid promoter – GFP] (LacI+)
2. pBR322 TetR + [LacI hybrid promoter – GFP] (lacI-)
3. pTrc99A + [placQI – GFP] (placQI is constitutive promoter)
4. pTrc99A + [LacI promoter – GFP]
5. pBR322 TetR + [LacI promoter – GFP]

Repressor LacI expression:
pTrc99A expresses LacI
pBR322 TetR does NOT express LacI

Antibiotics resistance:
pTrc99A gives ampicillin-resistance
pBR322 TetR gives ampicillin-resistance
[LacI hybrid promoter – GFP] gives kanamicin-resistance
[placQI – GFP] gives kanamicin-resistance

Procedure:

AHL assay Standard protocol