Team:Linkoping Sweden/Results

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<h2 class="clickable ">Biobricks</h2>
<h2 class="clickable ">Biobricks</h2>
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<p>These discussed combinations will result in 4 different biobricks which are the two combinations of RFP linked to either epitope 2(Fig.1) or all 5 epitopes (Fig.2) as well as the two combinations of MCherry with either epitope 2(Fig.1) or all 5 epitopes (Fig2).  The protein expressed from the biobrick including epitope 2 linked to RFP/MCherry will thus be used in combination with the monoclonal antibodies which are specific for epitope 2 as mentioned previously and the 5 different epitopes linked to RFP/MCherry will be used in combination with the polyclonal antibodies. Worth mentioning as well is that all the 4 biobricks will contain a sequence for Promotor + RBS as well as a sequence for His-TEV.</p>
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<p>These discussed combinations will result in 4 different biobricks which are the two combinations of RFP linked to either epitope 2 (Fig.1) or all 5 epitopes (Fig.2) as well as the two combinations of MCherry with either epitope 2 (Fig.1) or all 5 epitopes (Fig.2).  The protein expressed from the biobrick including epitope 2 linked to RFP/MCherry will thus be used in combination with the monoclonal antibodies which are specific for epitope 2 as mentioned previously and the 5 different epitopes linked to RFP/MCherry will be used in combination with the polyclonal antibodies. Worth mentioning as well is that all the 4 biobricks will contain a sequence for Promotor + RBS as well as a sequence for His-TEV.</p>
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Revision as of 19:29, 15 October 2014

Our vision is to create a biobrick including the sequence of the Ara h1 protein linked to a red fluorescent protein so that we in turn can express the protein and thus provide an interaction of this protein complex with the antibodies. To ensure ourselves that this epitope-red fluorescent protein complex will bind to the Ara h1 specific IgG antibodies several ideas of biobrick-setup was brought to mind. Since we use both monoclonal antibodies specific for epitope 2 of Ara h1 and polyclonal antibodies specific for several epitopes of 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) of Ara h1 linked to a red fluorescent protein. However, since there are two different mutants of the red fluorescent protein (called RFP and MCherry) we decided to create two setups of every biobrick combination to ensure that the best possible detection by FRET is used since RFP and MCherry slightly differs in their wavelength areas. The main idea is to practically test this FRET effect in both epitope-RFP and epitope-MCherry combinations by fluorescence which hopefully will prove to us which mutant is best suited to use and thus prove that our theory actually works.

Linköping University
581 83 Linköping, Sweden
liuigemgroup@gmail.com
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