Difference between revisions of "Part:BBa K530008:Experience"
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== '''Team Kyoto 2017''' ==
 
== '''Team Kyoto 2017''' ==
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<i>S. cerevisiae</i> expressing GFP from this TDH3 promoter was cultured and observed with a fluorescence microscope. The strong green fluorescence was observed. (fig 1 )
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<i>S. cerevisiae</i> expressing GFP from this TDH3 promoter was cultured and observed with a fluorescence microscope.<br>The strong green fluorescence was observed. (fig 1 )
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<html>
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''[https://static.igem.org/mediawiki/2017/4/48/GFP_green.jpg]'''
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<figure>
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Figure1: Observation of <i>S. cerevisiae</i> expressing EGFP by this TDH3 promoter with a fluorescence microscope.
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<img src="https://static.igem.org/mediawiki/2017/4/48/GFP_green.jpg" alt="Yeast expressing EGFP" style="float:center;width:350px;height:250px;">
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        </figure>
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Figure1: Observation of <i>S. cerevisiae</i> expressing EGFP from this TDH3 promoter with a fluorescence microscope.
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<br>
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<br>
 
<i>B. xylophilus</i> was cultured using this green fluorescent yeast as a bait. After several days, confocal observation confirmed that the intestine of <i>B. xylophilus</i> was filled with EGFP.  (fig 2 )  
 
<i>B. xylophilus</i> was cultured using this green fluorescent yeast as a bait. After several days, confocal observation confirmed that the intestine of <i>B. xylophilus</i> was filled with EGFP.  (fig 2 )  
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<figure>
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[[File:アフリカツメガエル結果injection.png|500px|thumb|center| '''Figure2 : <i>B. xylophilus</i> full of EGFP in its esophagus.
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<img src="https://static.igem.org/mediawiki/parts/8/82/B._xEGFP.jpg" alt="<i>B. xylophilus</i> filled with EGFP in its intestine" style="float:center;width:350px;height:250px;">
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        </figure>
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''']]
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Figure2 : <i>B. xylophilus</i> full of EGFP in its intestine.
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</html>
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This means that using the TDH3 promoter (BBa_K530008) makes it possible to express a very large amount of EGFP in <i>S. cerevisiae</i>, and that its expression level is sufficient to trace the intestinal tract of nematodes feeding on yeast. The above results indicate that ''' [https://parts.igem.org/Part:BBa_K530008  BBa_K530008] ''' and ''' [https://parts.igem.org/Part:BBa_K1875003  BBa_K1875003] ''' work effectively as a new usage that has not been attempted so far, greatly extending the function of each part.
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<br>
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<br>
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This means that using the TDH3 promoter (BBa_K530008) makes it possible to express a very large amount of EGFP in <i>S. cerevisiae</i>, and that its expression level is sufficient to trace the intestinal tract of nematodes feeding on yeast.
 +
<br>The above results indicate that ''' [https://parts.igem.org/Part:BBa_K530008  BBa_K530008] ''' and ''' [https://parts.igem.org/Part:BBa_K1875003  BBa_K1875003] ''' work effectively as a new usage that has not been attempted so far, greatly extending the function of each part.
  
 
===User Reviews===
 
===User Reviews===
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<!-- End of the user review template -->
 
<!-- End of the user review template -->
 
<!-- DON'T DELETE --><partinfo>BBa_K530008 EndReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K530008 EndReviews</partinfo>
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===NAU-CHINA 2017 iGEM===
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[[File:Tdh3.jpeg|500px|center]]
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Figure. Fluorescence intensity of different colonies of TDH3 promoter and improved TDH3 promoter. 
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We construct the device：
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TDH3 promoter+GFP+ADH1 terminator
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TDH3 promoter is a truncated promoter, GFP is a mutant. These is a invariable sequence upstream of GFP coding sequence.
 +
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The responses of promoter strengths of artificial TDH3 promoter and native promoter TDH3 were comprehensively compared. The strength of improved TDH3 was always higher than native promoter TDH3，but varied along with the genetic background of host.
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===TDH3 from kit plate===
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[[File:TDH3.jpg|600px|center]]
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TDH3 from kit plate,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly. It indicates this part could work.
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[[File:Yeast tutu.png|600px|center]]
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Truncated TDH3 promoter,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly.

Latest revision as of 02:25, 2 November 2017


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K530008

Team Kyoto 2017

S. cerevisiae expressing GFP from this TDH3 promoter was cultured and observed with a fluorescence microscope.
The strong green fluorescence was observed. (fig 1 )

Yeast expressing EGFP
Figure1: Observation of S. cerevisiae expressing EGFP from this TDH3 promoter with a fluorescence microscope.

B. xylophilus was cultured using this green fluorescent yeast as a bait. After several days, confocal observation confirmed that the intestine of B. xylophilus was filled with EGFP. (fig 2 )
<i>B. xylophilus</i> filled with EGFP in its intestine
Figure2 : B. xylophilus full of EGFP in its intestine.

This means that using the TDH3 promoter (BBa_K530008) makes it possible to express a very large amount of EGFP in S. cerevisiae, and that its expression level is sufficient to trace the intestinal tract of nematodes feeding on yeast.
The above results indicate that BBa_K530008 and BBa_K1875003 work effectively as a new usage that has not been attempted so far, greatly extending the function of each part.

User Reviews

UNIQe4b1903d3f4c0b3f-partinfo-00000001-QINU UNIQe4b1903d3f4c0b3f-partinfo-00000002-QINU

NAU-CHINA 2017 iGEM

Tdh3.jpeg

Figure. Fluorescence intensity of different colonies of TDH3 promoter and improved TDH3 promoter.

We construct the device： TDH3 promoter+GFP+ADH1 terminator TDH3 promoter is a truncated promoter, GFP is a mutant. These is a invariable sequence upstream of GFP coding sequence.

The responses of promoter strengths of artificial TDH3 promoter and native promoter TDH3 were comprehensively compared. The strength of improved TDH3 was always higher than native promoter TDH3，but varied along with the genetic background of host.

TDH3 from kit plate

TDH3.jpg

TDH3 from kit plate,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly. It indicates this part could work.

Yeast tutu.png

Truncated TDH3 promoter,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly.