Difference between revisions of "Part:BBa K2271105:Experience"
 
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__NOTOC__
 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
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===Applications of BBa_K2271105===
 
===Applications of BBa_K2271105===
  
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<h1>Import of the fluorescent protein mTurquoise</h1>
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====Import of fluorescent proteins into the peroxisomal matrix====
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<h2>PEX5 variant with wildtype PTS1</h2>
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https://static.igem.org/mediawiki/2017/b/b0/Artico_15wtp13.png
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<h2>PEX5 variant with PTS1 variants</h2>
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<h3> PTS1 variant P2 </h3>
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[[Image:Artico_15wtp13.png|frame|center|'''Figure 1:'''PEX5 knock out strain, transformed with the R15 variant, mTurqouise tagged with the PTS1 and the PEX13 membrane anchor fused to mRuby.]]
  
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https://static.igem.org/mediawiki/2017/a/ae/Artico_15p3p13.png
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[[Image:Artico_15p3p13.png|frame|center|'''Figure 2:'''PEX5 knock out strain, transformed with the R15 variant, mTurqouise tagged with the PTS1* P3 and the PEX13 membrane anchor fused to mRuby.]]
  
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  The figures 1 and 2 show two different experiments. The first in figure 1 is the control if the PEX5 variant R15 imports proteins tagged with the natural PTS1 and the second in figure 2 is an actual experiment if the variant imports proteins tagged with one of our new PTS1* variants &minus; in this case it was P3 (-YTNQE). One can see that there is no import in both cases. While PEX13-mRuby marks the peroxisomal membrane, mTurqouise is located in the cytosol &minus; this indicates that there is no import.
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  We wanted to make a screening for functional PTS1* variants but unfortuneately the Violacein assay planned for that (see our [http://2017.igem.org/Team:Cologne-Duesseldorf/Design#h3-3 wiki article] for more information) did not work out. As outlook we propose to find a fitting PTS1* hence the molecular dynamics simulations showed promising results.
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===User Reviews===
 
===User Reviews===
 
<!-- DON'T DELETE --><partinfo>BBa_K2271105 StartReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K2271105 StartReviews</partinfo>

Latest revision as of 10:17, 29 October 2017

Applications of BBa_K2271105

Import of fluorescent proteins into the peroxisomal matrix

Figure 1:PEX5 knock out strain, transformed with the R15 variant, mTurqouise tagged with the PTS1 and the PEX13 membrane anchor fused to mRuby.
Figure 2:PEX5 knock out strain, transformed with the R15 variant, mTurqouise tagged with the PTS1* P3 and the PEX13 membrane anchor fused to mRuby.

The figures 1 and 2 show two different experiments. The first in figure 1 is the control if the PEX5 variant R15 imports proteins tagged with the natural PTS1 and the second in figure 2 is an actual experiment if the variant imports proteins tagged with one of our new PTS1* variants − in this case it was P3 (-YTNQE). One can see that there is no import in both cases. While PEX13-mRuby marks the peroxisomal membrane, mTurqouise is located in the cytosol − this indicates that there is no import.
We wanted to make a screening for functional PTS1* variants but unfortuneately the Violacein assay planned for that (see our [http://2017.igem.org/Team:Cologne-Duesseldorf/Design#h3-3 wiki article] for more information) did not work out. As outlook we propose to find a fitting PTS1* hence the molecular dynamics simulations showed promising results.


User Reviews

UNIQd23f4d046722112d-partinfo-00000000-QINU UNIQd23f4d046722112d-partinfo-00000001-QINU