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Team:NRP-UEA-Norwich/Week3
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Welcome to the NRP UEA iGEM 2012 Wiki Lab Book
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Contents |
Over the weekend the group met up to discuss the two potential projects we could go forward with using our synthesised gene and the Biobrick it will ultimately produced. We developed two main themes; one was a multi-sensor system sensing oxygen-containing physiologically-relevant molecules and reporting on the levels of each molecule; the other was a comparator circuit sensing levels of molecular species and using novel gene expression to express reporters for specific molecules within the environment to quantitatively measure them. We decided on the latter as we felt it would have the highest scientific impact, a diagram of the gene system for one of the sensors is given below.
The concept behind the project involves the mRNA control region of one sensor binding to the mRNA control region of a second sensor and thus for one sensor to 'switch off' the other to a certain degree, and for the overhang to be used to measure levels of a substrate molecule. For example in our nitric oxide-sensing project a sensor for nitric oxide and nitrites (sensor 1) may be used in conjunction with a sensor for only nitrites (sensor 2); the control region of mRNA produced from sensor 2 would switch off sensor 1 per the levels of nitrite in the environment, and therefore all expression of sensor 1 would be directly related to nitric oxide levels in the atmosphere.
Amy (our artist) came to visit us today in order to discuss our video. She, Joy and Rachel began to storyboard the film and work out how to incorporate the decision made over the weekend into the video. They came up with some fantastic ideas and the whole team sat down to discuss them before Amy left to organise making props. Joy also got in touch with the UEA Drama School in order to see whether we could hire a studio in order to film our video.
From the transformation of the two promoters carried out on Friday, we inoculated some colonies into culture. This will be used later for characterisation of the promoter. From the two plates, we noticed that one plate had bigger colonies and one had very small colonies. The larger colonies were from the agar plate inoculated with transformed E.coli with promoter 1.
To characterise the promoters and other Bio-Bricks we may be interested in, an input of NO will be needed. Our solution to this is to use a nitrate salt like potassium nitrate. A stock solution (1M) was made up today by Lukas. This was taken to be autoclaved.
We were going to inoculate the E.coli with the transformed Pyear promoters that
After our hunt for the fluorometers and speaking to Dr. Yeoman the previous day we decided that we needed to find whether we could use fluorescent proteins today in order to go on to design the DNA for the control region and reporter/enzyme. After a social media call-out for fluorometers in our school we were pointed in the direction
