Part:BBa_K1655002

Amphiphilic proteins can be used to form intracellular micelles or vesicles.

Figure 1. Amphiphilic Brick

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 that the size of this insert in the pSB1C3 backbone is correct. See figure 2 for a gel electrophoresis image. DNA from the colony which produced the product seen in figure 10, 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 figure 3 where the last two wells before the ladder show a correct plasmid size. Although the sequencing results are unclear, the correct sized mRNA seemed to be produced. This lead us to believe, that the Amphiphilic protein was indeed transcribed and possibly translated as well. We thought that we weren't able to detect the micelles and vesicles because of low accuracy of the microscopic images or because our expression level was too high. Our promoter was 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.

Note! 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 2). 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 blue chromoprotein.

Click the images to enlarge them

Figure 2. Amphiphilic insert on gel

Figure 3. GFP-Amphiphilic fusion on gel

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
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 13
  
• 1000
  COMPATIBLE WITH RFC[1000]