Achievements

1. We registered our Team.
2. We completed the judging form.
3. We designed a Team Wiki.
4. We will present a poster and a talk at the iGEM Jamboree.
5. We documented several new biobricks for the 2013 iGEM competition, including K1166000, K1166001, K1166003, K11660004, K1166005. We also implement the biobrick K905000, and reported our results in their “experience page” at the registry.
6. We experimentally validated several of our new biobricks K1166000, K1166001, K1166003, K1166004, K1166005 and even our new device “K1166002”; and its documentation is shown in the “Main page” section of our parts and device registry page.
7. We submitted our parts and device to the Registry in time.
8. We explained several considerations we took into account while designing our project in order to address the security and safety issues concerning the use of therapeutic proteins and their handling; they are all presented in the “Project” tab of our webpage.
9. We designed our device (K1166002) thinking in the improvement of several existing biobricks all at once, the biobricks taken into account were AraC/pBAD:BBa_I0500, HlyA: BBa_K554002, HlyB: BBa_K554007 and HlyD: BBa_K554008. We made sure to add some information to each of their experience page in the registry, and created its own for our device.
10. This year we managed to collaborate with two iGEM teams: Team Buenos_Aires, and Team UC_Chile. For team Buenos_Aires, we manage to characterize their part BBa_K1106003 consisting of an mRFP generator under the Ars Promoter, all our characterization information can be found at our results Tab. For Team UC_Chile we helped them increase the outreach of their video game in our country, being visited by several of our wiki, and facebook followers.
11. Taking into consideration several implications regarding ownership and sharing of iGEM projects, we decided to develop a guide, novel to iGEM, referring to patenting issues in our country and an useful tool which many teams can use to understand more about the commercial value of their project, if they wish to continue working on it, or even develop a starter project. We’ve provided some guidelines and the link to our very own “QuickLook” in the “Project” tab of our webpage.

Collaboration

One of the most important parts of science and scientific progress is commonly forgotten by researchers. Working and communication between teams is not only beneficial for both in technical terms, but also opens a bridge that is essential for science; collaboration.

This year we crossed words with a fellow Latin American team. Team: Buenos_Aires started the conversation and our ideas began to flow we talked about how we were achieving hypoxia for our hypoxic promoters characterization and how they were measuring their arsenic sensitive promoters. After some discussion we concluded that we could improve each other’s measurements and agreed to make the collaboration.

When we received the part Team: Buenos_Aires we proceeded with transforming its DNA into Escherichia coli TOP10 chemocompetent cells. Selection of transformants was done with Tetracicline and we continued with a restriction analysis using EcoRI and PstI enzymes. The resulting agarose gel is shown by duplicate in the following pictures.

After transformants were proven to have the correct plasmid by restriction analysis, we started induction with sodium arsenite. Different concentrations ranging from 0 ppb to 1000ppb were used as induction and measurements were done at 1, 2, 4, 6, 8 hours.

In each column, 10mL cultures of E.coli transformed with the measurement device were grown until 0.5 OD600. After that, cultures were induced with different concentrations (in ppb) of sodium arsenite. Measurements of RFP fluorescence (AU) and OD600 were taken every time specified in the time (h) column and AU/OD600 is reported. A public water source and water from a nearby lake were also assayed.

Using the last measured fluoresence/OD600 value plotted against the concentration of arsenite used as inductor, a calibration curve was obtained.

With the use of the newly obtained calibration curve we also measured two samples that we obtained from our neighborhood. We took samples from a nearby lake and from a public water source. Following the same procedure, we grew E.coli cells until .5 OD600 and induced with the samples we obtained (after using a 0.0002 mm syringe microfilter) until 8 hours of induction. After measuring AU/OD600 for 8 hours we obtained the following data:

Taking the latter into account we might be more careful from where do we have contact with water. These results encourage us to inform the responsible authorities in the future about the current status of the water and have the samples analyzed by an already established method.