Team:UNAM-Genomics Mexico/Modules/In vivo

Coupling together Biological Chassis

In order to enable the light based communication between bacteria, we have designed a tertiary cycle of different bacteria chassis, assembling each module of reception and emission to construct a light-based feedback loop of red-green-blue light, which will make the proof of concept of communication over distance and proper signal decoding.

First Chassis

Input: Red light

Output: Green light

First Chassis: Red Light reception module coupled to Green Light emitter module.

Reception module: Blue Light Photosensor LovTAP

Working in this chassis, we decided to use as blue light receptors: LovTAP photosensor and the native blue light inducible promoter from E.coli.

OBJECTIVES:

• Construct an improved version of the LovTAP photosensor device under the regulation of different constitutive promoters, due to the previous version of LovTAP designed by 2009 Lausanne team didn´t have a differential behavior under the inverting regulator sensitive to LacI and CAP protein, their results showed that the expression levels of LovTAP didn’t show differences to the induction with IPTG. As well, we included a punctual mutation to change the ILE427 by a PHE427, as was proposed by the model results of the team iGEM09_EPF-Lausanne. With this mutation LovTAP should react faster and the conformational change should be more stable (the protein stays in the active form for longer, under light induction).

• Characterize LovTAP reporter systems, both repressor and activator activity.

The structure of the device is designed as follows:

Second Chassis

Input: Green light

Output: Blue light

Second Chassis: Green Light reception module coupled to Blue Light emitter module.

Third Chassis

Input: Blue light

Output: Red Light light

Third Chassis: Blue Light reception module coupled to Red Light emitter module.

Tertiary cycle

With the previous chassis assembled, the interchange and processing of information go through an activation signaling cascade, that is illustrated in the next image:

Coupling together Biological Chassis through Light communication