Difference between revisions of "Team:Exeter/Parts"
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<h6>Description</h6> | <h6>Description</h6> | ||
| − | <p id="pp">KillerRed is a red fluorescent protein that generates reactive oxygen species after illumination with light between the wavelengths of 540- | + | <p id="pp">KillerRed is a red fluorescent protein that generates reactive oxygen species after illumination with light between the wavelengths of 540-580 nm[1].</p> |
<p id="pp">We further characterised this kill switch by illuminating induced cultures 24 hours after induction with IPTG as well as uninduced cultures. After 6 hours in the light box CFU’s were counted to determine if the kill switch was successful. This was also reproduced on cultures grown in a ministat for 120 and 168 hours to test how long the kill switch remains functional. The full protocol can be found <a href="https://2016.igem.org/Team:Exeter/Project#section_4">here</a> and the results can be found <a href="https://2016.igem.org/Team:Exeter/Project#section_2">here</a>. </p> | <p id="pp">We further characterised this kill switch by illuminating induced cultures 24 hours after induction with IPTG as well as uninduced cultures. After 6 hours in the light box CFU’s were counted to determine if the kill switch was successful. This was also reproduced on cultures grown in a ministat for 120 and 168 hours to test how long the kill switch remains functional. The full protocol can be found <a href="https://2016.igem.org/Team:Exeter/Project#section_4">here</a> and the results can be found <a href="https://2016.igem.org/Team:Exeter/Project#section_2">here</a>. </p> | ||
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<p id="pp">KillerOrange has an excitation maximum of 512 nm and emission maximum at 555 nm and its absorbance spectrum has two peaks, at 455 and 514 nm. <p> | <p id="pp">KillerOrange has an excitation maximum of 512 nm and emission maximum at 555 nm and its absorbance spectrum has two peaks, at 455 and 514 nm. <p> | ||
| − | <p id="pp"> | + | <p id="pp">The mechanism by which KillerOrange kills cells isn’t fully understood yet. However, it is believed that KillerOrange's ability to generate reactive oxygen species depends on a water-filler channel reaching the chromophore area from the end cap of the ß-barrel [3].</p> |
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<p id="pp">We characterised this part in the same way as KillerRed (<a href="https://2016.igem.org/Team:Exeter/Project#section_4">protocol</a>), and the results of which can be found <a href="https://2016.igem.org/Team:Exeter/Project#section_2">here</a></p> | <p id="pp">We characterised this part in the same way as KillerRed (<a href="https://2016.igem.org/Team:Exeter/Project#section_4">protocol</a>), and the results of which can be found <a href="https://2016.igem.org/Team:Exeter/Project#section_2">here</a></p> | ||
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<div id="section_3" class="link_fix"></div> | <div id="section_3" class="link_fix"></div> | ||
<div id="contentTitle"> | <div id="contentTitle"> | ||
| − | + | Lysozyme | |
</div> | </div> | ||
<h6>Name:</h6> | <h6>Name:</h6> | ||
| − | <p id="pp">pT7 Lysozyme | + | <p id="pp">pT7 Lysozyme E.coli codon optimised, signal peptide, flag tag</p> |
<h6>Description:</h6> | <h6>Description:</h6> | ||
| − | <p id="pp">Lysozyme | + | <p id="pp">Lysozyme from chicken egg white, attacks the cell wall in bacteria by hydrolysing the ß-1,4 linkages between N-acetylmuramic acid and N-acetylglucosamine of peptidoglycan [4].</p> |
<p id="pp"> </p> | <p id="pp"> </p> | ||
<p id="pp">Characterisation of this part involved using the Enzcheck assay kit which detects lysozyme activity as a measure of fluorescence as well as using the same protocol we used to characterise KillerRed (<a href="https://2016.igem.org/Team:Exeter/Project#section_4">protocol</a>)and the results of which can be found <a href="https://2016.igem.org/Team:Exeter/Project#section_2">here</a></p> | <p id="pp">Characterisation of this part involved using the Enzcheck assay kit which detects lysozyme activity as a measure of fluorescence as well as using the same protocol we used to characterise KillerRed (<a href="https://2016.igem.org/Team:Exeter/Project#section_4">protocol</a>)and the results of which can be found <a href="https://2016.igem.org/Team:Exeter/Project#section_2">here</a></p> | ||
| − | <p id="pp">Here we are submitting | + | <p id="pp">Here we are submitting lysozyme as a composite part under a T7 promoter (BBa_I712074), an Elowitz ribosome binding site (BBa_B0034) and a double terminator (BBa_B0015).</p> |
| − | <p id="pp">We have codon optimised the protein coding region, added a FLAG tag and signal peptide directing Lysozyme to the periplasm.The sequence for | + | <p id="pp">We have codon optimised the protein coding region, added a FLAG tag and signal peptide directing Lysozyme to the periplasm.The sequence for Lysozyme protein coding region can be found here (BBa_K1914004)</p> |
<h6> Biobrick Code </h6> | <h6> Biobrick Code </h6> | ||
Revision as of 11:13, 18 October 2016
