Team:UNICAMP-Brazil/Coliguard/Results

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The Coliguard - Results

Test and characterization of device BBa_K284022

We concluded the construction of the device BBa_K284022.

We confirmed and characterized the device. The results are shown next:

New biobrick.JPG

We confirm the construction through PCR and restriction analysis. Click here for details.

After the confirmation of the construction we characterized the part through the test that is described in the Notebook. Click here for details.

We confirmed that our part really works as expected!!!!

The OD measurements showed a decrease on the growth of induced strains transformed with BBa_K284022, in comparsion with induced strains transformed with BBa_K112806 (without the promoter) and the non-induced ones. The lysis of the cells expressing T4 endolysin can be visualized in the picture below. A graphic comparing the growing of induced and non-induced cultures after the induction is also shown.

Cell-lysis.png


Graph 1. Growth cellular after ITPG induction1.JPG

Besides that, the death of the induced cells capable of expressing the endolysin was confirmed by the absence of growing colonies on the plates. On the other hand, the cultures that weren't expressing the endolysin grew normally.

Non-induceds.png
Induced.png

It was also possible to observe the overexpression of T4 endolysin through an SDS-PAGE.

SDS-PAGE.png
Legenda.png


Cre-Recombinase without ATG's biobrick assemble

The biobrick of Cre-Recombinase without the ATG start codon was successful assembled. You can check it on registry catalog under accession name BBa_K284031. This biobrick was also the only one we constructed according to the standard assembly strategy.

The confirmation of it's assemble can be checked on our notebook, on October 9th and October 10th.

Although we assembled this part’s biobrick, we didn’t have enough time to assemble the entire device, i.e., the construction with the AGTC repetition and the constitutive promoter. Therefore, we didn’t performed any experiments in order to characterize this part.