Difference between revisions of "Team:Aalto-Helsinki/Parts"
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<figcaption><center><b>Figure 9.</b> Amphiphilic Brick</center></figcaption> | <figcaption><center><b>Figure 9.</b> Amphiphilic Brick</center></figcaption> | ||
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+ | <p>We submitted a sole amphiphilic brick, containing only the coding region of the ampihphilic protein. This brick can be used to produce intracellular micelles or vesicles under any chosen expression system. Our amphiphilic protein is constructed so that the hydrophilic part is translated first. According to Huber et al. this allows for the formation on of intracellular micelles and vesicles. If the hydrophobic domain was to be translated first, the amphiphils would only form micelles.<p> | ||
− | + | <p><b>Validation:</b> Our amphiphilic brick's sequencing results were unclear, as the brick is mainly built of short repeats. We have however been able to show the size on this insert in the pSB1C3 backbone is the correct. See figure 10 for a gel electrophoresis image. The DNA from the colony which produced the product seen in well 2 was sent to the registry. We were also able to show, that once this Amphiphilic brick is fused to a GFP, the GFP is expressed and again, the construct size is correct. See <span style="color:red">figure 11.</span> Although the sequencing results are unclear, there seemed to be no premature stop codons in the sequence. This leads us to believe, that the Amphiphilic protein was indeed translated. We weren't able to detect the micelles and vesicles because of low accuracy of the microscopic images or because our expression lever was too high Our promoter is 10 times more efficient than the one that has been previously used to produce micelles and vesicles with this amphiphilic protein. The high concentration of amphiphils is likely to interfere with the micelle and vesicle formation.<br/><br/></p> | |
− | <p><b>Validation:</b> Our amphiphilic brick's sequencing results were unclear, as the brick is mainly built of short repeats. <span style="color:red"> | + | |
− | <p style="color:red">The | + | <p><span style="color:red"><b>Note!</b></span> The submitted BioBrick does most likely not contain the correct sequence. Restriction analysis lead us to initially believe we had the correct insert, as the insert size in pSB1C3 backbone matched that of the correct sequence (figure 1). However, as we proceeded to the sequencing results of the then already submitted brick a day before the wiki freeze, we found out that the sequence was not that of the amphiphilic protein, but instead that of the <a href="http://parts.igem.org/Part:BBa_K592009">blue chromoprotein</a>.</p> |
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Revision as of 19:51, 17 September 2015