Difference between revisions of "Part:BBa K1655002"

Revision as of 11:10, 17 September 2015

Amphiphilic proteins can be used to form intracellular micelles or vesicles. This is the coding sequence for an amphiphilic protein. The protein has both a hydrophilic and a hydrophobic domain, each about 300bps in length. In this brick, the hydrophilic domain is translated first. According to Huber et al. this allows for the formation of intracellular vesicles rather than micelles. The coding sequence is derived from this article.

Usage and Biology

Validation: 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 1 for a gel electrophoresis image. 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 figure 2. 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. Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 616
23
INCOMPATIBLE WITH RFC[23]
Unknown
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 13
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K1655002 short</partinfo>
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 This is the coding sequence for an amphiphilic protein. The protein has both a hydrophilic and a hydrophobic domain, each about 300bps in length. In this brick, the hydrophilic domain is translated first. According to <a href="http://www.nature.com/nmat/journal/v14/n1/abs/nmat4118.html">Huber </a><i>et al.</i> this allows for the formation of intracellular vesicles rather than micelles. The coding sequence is derived from this article.
+</html>
-<!-- Add more about the biology of this part here
+<!-- Add more about the biology of this part here-->
 ===Usage and Biology===
+<html><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 <span style="color:red">figure 1</span> for a gel electrophoresis image. 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 2.</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.</html>
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 <span class='h3bb'>Sequence and Features</span>