Difference between revisions of "Team:Cardiff Wales/results"

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<p>This image shows a protein gel that team TSH created to test whether part <a href="http://parts.igem.org/Part:BBa_K2404016"> BBa_K2404016 </a> had been expressed in <i> N. benthamiana </i>. There was no band present where we anticipated, so concluded that it had not worked.<br><br><br><br>
 
<p>This image shows a protein gel that team TSH created to test whether part <a href="http://parts.igem.org/Part:BBa_K2404016"> BBa_K2404016 </a> had been expressed in <i> N. benthamiana </i>. There was no band present where we anticipated, so concluded that it had not worked.<br><br><br><br>
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<center> <h3>Team Plant Promoter</h3><br><br>
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<p> Team plant promoter were responsible for creating any constructs containing TSHH or Luc+ under the control of one of our 4 inducible promoters. They needed to isolate the promoters from <i> Arabidopsis </i> DNA, amplify them using PCR, and extract them, then put them into level 0 parts. Following that, they created level 1 constructs with these promoters, containing either TSHH or Luc+ as the CDS, and usually with the NOS terminator from <i> Agrobacterium tumefaciens </i>, but some of the final constructs somehow lacked these as shown on the <a href"https://2017.igem.org/Team:Cardiff_Wales/compositeparts"> composite parts </a> page. Some of their results are shown below.<br><br></p>
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Revision as of 13:32, 29 October 2017




Our Results




During our project, we had three smaller teams within our main team. Each of these teams had different but interlinked objectives, and consequently resulted in different sets of results. The results of each team are shown with descriptions on this page.




Team Luciferase



Team luficerase created the 35S:Luc+:NosT construct , and quantified the 35S promoter using a luciferase reporter assay.



This photo shows the DNA gel that showed the level 1 reaction to create the 35S:Luc+:NosT construct had worked. The DNA solution that this was taken from was later sequenced to confirm that the construct was correct.





This image shows leaf discs taken from plants infiltrated with Agrobacterium containing the 35S:Luc+:NosT construct. The red regions show high levels of luciferase expresion.





Team TSH



Team TSH was responsible for creating constructs containing TSH or TSHH (TSH antagonist with His-tags for purification), controlled by the LexA promoter. This is part BBa_K2404016 . They also attempted to create a part with 35S:TSHH:NosT, but these ligations failed. Using their TSHH constructs, they infiltrated N. benthamiana and performed a protein extract which was ran on a protein gel to identify the presence of the antagonist.



This photo shows the DNA gel that showed the level 1 reaction to create the LexA:TSHH:NosT construct had worked. The DNA solution that this was taken from was later sequenced to confirm that the construct was correct.





This image shows a protein gel that team TSH created to test whether part BBa_K2404016 had been expressed in N. benthamiana . There was no band present where we anticipated, so concluded that it had not worked.





Team Plant Promoter



Team plant promoter were responsible for creating any constructs containing TSHH or Luc+ under the control of one of our 4 inducible promoters. They needed to isolate the promoters from Arabidopsis DNA, amplify them using PCR, and extract them, then put them into level 0 parts. Following that, they created level 1 constructs with these promoters, containing either TSHH or Luc+ as the CDS, and usually with the NOS terminator from Agrobacterium tumefaciens , but some of the final constructs somehow lacked these as shown on the composite parts page. Some of their results are shown below.