Team:IPN-UNAM-Mexico/Parts

Biobricks

[[Image:Month-icon.png | 50px]]Biobricks
We contribute to the registry with 11 new biobricks, 5 of them are favorite biobricks from the team and in this section we document them:

Projects
These two biobricks are the main Activator-Inhibitor system, as showned below they are in two different plasmids, and the idea is transform Top10 cells with them in petri dishes with IPTG and ATC.

Auxiliary biobricks
This biobricks are though like construction intermediates necessary to build the main and project biobricks.

[[Image:Month-icon.png | 50px]]Plasmids
Check registry for more info with the name of the biobrick

[[Image:Month-icon.png | 50px]]Results
As shown as follow we were able to test the viability of the Activator module coupled to its Regulatory module.

The activator and regulatory modules are composed of the following BioBricks: BBa_K266006, BBa_K266004, you can check their full description on the Modules section on the top of this page.

This BioBricks were into the vectors PSB1C3 y PSB1A2, with resistence to Chloramphenicol and Ampiciline respectively. The plasmids were transformed into an E. coli TOP10 strain and they were plated in agar plates containing Chloramphenicol and Amipiciline. After 24 hours at 37°C they were observed into an epifluorescence microscope Zeiss. In one plate 160mM of IPTG was previously added to the agar pate and in the other no IPTG was added.

The following pictures were taken at the optimal exitation wavelenght of the GFP (475nm) under the same exposure and amplification conditions.

e basal production of GFP that can be seen in the following pictures in basal conditions inticates that there is transcription of the BioBrick BBa_K266006. From this we can infer that that there is a basal production of LasI and thus of PAI, one of the requirements we need for our design to work properly.

As can be appreciated in the pictures with 160mM of IPTG there is an increase in the production of GFP, this indicates that the GFP was only basally expressed in the absence of IPTG, demonstrating that the Lac inversor was effectively repressing lasR, and there could be no induction in the production of GFP and LasI.

When the repression of LacI was released by adding IPTG to the media, LasR was produced and together with the basal production of PAI (by LasI) there was a positive feedback in the production of LasI, resulting in an increase of the production of both LasI and GFP (since they were under the control of the same promoter).

With this single experiment we were able to test the proper functioning of all the elements in these two modules.