Team:Linkoping Sweden/Results


Our vision is to create a Biobrick which includes the sequence of the Ara h1 protein linked to a red fluorescent protein. We want to accomplish this so that we in turn can express the protein and thus provide an interaction of this protein complex with the antibodies. To ensure that the epitope-red fluorescent protein complex will bind to the Ara h1 specific IgG antibodies several ideas for Biobrick-design was brought to mind. Since we use both monoclonal antibodies specific for epitope 2 on Ara h1 and polyclonal antibodies specific for several epitopes on Ara h1 we decided to create a Biobrick consisting of epitope 2 of Ara h1 linked to a red fluorescent protein as well as a biobrick consisting of five wisely chosen epitopes (epitope 22, epitope 1, epitope 3, epitope 4, and epitope 17) from Ara h1 linked to a red fluorescent protein. However, since there are two different mutants of red fluorescent protein (called RFP and MCherry respectively) we decided to create two setups of every Biobrick combination to ensure that the best possible detection by FRET is attained, as RFP and MCherry differ slightly in their excitation and emission wavelengths. The main idea is to practically test this FRET effect in both epitope-RFP and epitope-MCherry combinations by fluorescence. The data from these tests will show us which mutant is best suited to use and thus prove that our theory works.

Linköping University
581 83 Linköping, Sweden
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