The aim of our experiments is to demonstrate the coloration, for that we have done different experiments:

The goal is to introduce the constructions (pcst-RBS-CinR) and (pCin-lycopene) on the same bacterium, the constructions are supported on pSB3C5 and pSB1A2 respectively., by adding externel AHL molecule, this latter can enter in the bacterium and forms with CinR molecules complex that allows the activation of Lycopene production, after few times we can see the red color.

CLONING OF THE CONSTRUCTION pCst-RBS-CinR IN pSB3C5:

• Restriction using EcoRI and Pst1.
• Ligation: standard protocol was performed; Pcst-RBS-CinR was inserted into pSB3C5 plasmids
• Spreading over Petri dish.
• Better cloning rate (New stock of biobrick Miniprep)
• Do checking, PCR cheking.
• To confirm the result gel checking was performed.

pCin-Lycopene CONSTRUCTION IN pSB1A2 PREPARATION

• Culture overnight (the biobrick Miniprep was prepared).

Double transformation (introduction of the two plasmids contain pCst-RBS-CinR and pCin-Lycopene respectively).

• Spreading over Petri dish containing double resistances Ch and Amp.
• Good results, we choose 15 clones.
• PCR on clones was performed, 8 clones had the two plasmids.
• To confirm the result gel checking was performed.

PREPARATION OF THE 96 WELLS PLATE.

• We have tested our 8 clones with three different concentration of AHL (1mM, 0,5mM and 0,2mM).