Difference between revisions of "Part:BBa P10003:Experience"

 
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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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[http://2017.igem.org/Team:Cardiff_Wales Cardiff_Wales Team 2017]:
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how you used this part and how it worked out.
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We clearly demonstrate that leaf samples infiltrated with the 35S-OTMV:LUC+ construct show strong luciferase activity.<br> Therefore this construct is an excellent tool to act as a control for successful tobacco leaf infiltrations.
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Previous studies have shown that expression levels of infiltrated proteins can be very variable ([https://plantmethods.biomedcentral.com/articles/10.1186/s13007-015-0091-5 Bashandy <i>et al</i> 2015]). We also observe this as two leaves with the same treatments showed significant differences in expression (see below Z1 compared to Z2).
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https://static.igem.org/mediawiki/parts/e/e9/T--Cardiff_Wales--35S-LUC-7.png
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Our data show that expression levels do not coincide with the amounts of luciferin or agrobacterium we used in these experiments. Therefore in subsequent experiments we would use 0.5mM luciferin and an agrobacterium concentration of OD600 0.5.
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<img width=100% src="https://static.igem.org/mediawiki/parts/0/04/T--Cardiff_Wales--35S-LUC-12.png">
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<br><img width=100% src="https://static.igem.org/mediawiki/parts/6/6a/T--Cardiff_Wales--35S-LUC-11.png">
  
 
===Applications of BBa_P10003===
 
===Applications of BBa_P10003===

Revision as of 21:05, 30 October 2017


[http://2017.igem.org/Team:Cardiff_Wales Cardiff_Wales Team 2017]:

We clearly demonstrate that leaf samples infiltrated with the 35S-OTMV:LUC+ construct show strong luciferase activity.
Therefore this construct is an excellent tool to act as a control for successful tobacco leaf infiltrations.

Previous studies have shown that expression levels of infiltrated proteins can be very variable (Bashandy et al 2015). We also observe this as two leaves with the same treatments showed significant differences in expression (see below Z1 compared to Z2).

T--Cardiff_Wales--35S-LUC-7.png

Our data show that expression levels do not coincide with the amounts of luciferin or agrobacterium we used in these experiments. Therefore in subsequent experiments we would use 0.5mM luciferin and an agrobacterium concentration of OD600 0.5.

===Applications of BBa_P10003=== ===User Reviews=== BBa_P10003 StartReviews BBa_P10003 EndReviews