Team:Shasta Summit CA/Project

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 the team has and is 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

Doing some research the team was able to find an organism that was able to sense Carbon Monoxide and react. Using that gene they added that to the 1-9i plate, which contains an RBS and an RFP. This is going to be the reporter plasmid. Then they made a plasmid that would be the sensor. (The CooA gene.) Using the 3a assembly the team would combine the two plasmids onto the pSB1C3 plasmid backbone.

Testing The Bacteria

The team plans to expose the bacteria to Carbon Monoxide. They 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 they will remove the gas and add that to a cylinder filled with 1900ml of air. Then with that mixture they will pump that out into a container containing the bacteria. 100ml in a 2000ml total would make that 5% mixture.

3A Assembly

The team had a blast using the 3A assembly kit, learning all sorts of things about cutting plasmids and instituting their own. Although with some failures, the team moved on and created a successfully modified plasmid.