Our Project

We plan to ultimately create a bacteria that would be stimulated by the presence of CO to produce a red hue. The uses of this kind of organism/bacteria would include disaster relief in areas that do not have the access to the equipment needed to monitor the concentration of CO in the area. The goal would be to have the bacteria glow red in the presence of CO, and different colors based on the concentration. For example: It would glow a darker red and/or emit a smell if the level of CO in the air was at fatal levels. It is based upon the percentage of CO in the air. Carbon Monoxide is a fatal gas that is undetectable to humans. In iGEM we plan to use biobricks to make sensors. We plan to construct two sensors, one with the sensor and one with the reporter. The plasmid with the reporter with glow red if the levels of CO are at fatal levels. iGEM also requires outreach which we have and are currently doing; the outreach includes teaching and using a website to convey information.

The Plan

The Notebook

Every Tuesday we meet to make progress on our iGEM project. On Thursdays we go to the lab at skyline collage and do laboratory work, at the end of our meetings we discuss the progress we have made, and research new information for our next meeting!

Making the Bacteria

We will be using a two plasmid system. One plasmid will be a Sensor Plasmid and the other will be a

Sensor Plasmid

• Promoter (constitutive)

• RBS (Ribosomal Binding Site)

• CO Sensor Gene

• Terminator

Reporter Plasmid

• Operator for CO Sensor

• Promoter (inducible)

• RBS

• RFP Gene

• Terminator

Doing some research we were able to find an organism that was able to sense Carbon Monoxide and react. Using that gene we added that to the BBa_K516030 plate, which contains an RBS and an RFP. This is going to be the reporter plasmid. Then we made a plasmid that contains the sequence for the CooA protein, the sensor. Using the 3A assembly we will combine the two plasmids onto the pSB1C3 plasmid backbone.

Testing The Bacteria

We plan to expose the bacteria to Carbon Monoxide. We will start with a preliminary test of simply exposing the bacteria to carbon monoxide. The second set of tests they plan to do would be to create a gas mixture of 5% carbon monoxide, 1% carbon monoxide, and .1% carbon monoxide, respectively. Making sure that the mixture is correct is difficult, as measuring how much gas is release is difficult. So they filled a 2 liter graduated cylinder with water then pumped Carbon Monoxide into the container and stopped when there was approximately 100ml of water out of the graduated cylinder. With 100ml out that means there is 100ml of gas in the graduated cylinder. Then we will remove the gas and add that to a cylinder filled with 1900ml of air. Then with that mixture we will pump that out into a container containing the bacteria. 100ml in a 2000ml total would make that 5% mixture.

Results/Conclusions

Over the course of the year we have practiced with bacteria by making transformations and competent cells. Here is a diagram of the project we intend to complete:

Learning and Practicing

3A Assembly

We had a blast using the 3A assembly kit, learning all sorts of things about cutting plasmids and instituting their own. Although we made some mistakes, we were able to move on to successfully create a modified bacteria.