Difference between revisions of "Part:BBa K2078002:Experience"

 
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==Applications of BBa_K2078002==
  
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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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===Experience of CLSB-UK-2016 iGEM team with [[Part:BBa_K2078002]]===
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how you used this part and how it worked out.
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====Experimental Design====
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This consensus promoter was assembled with the reporter protein, blue-purple chromoprotein AmilCP. The construct was carried on the pSB1C3 plasmid for expression and amplification in E-Coli. The plasmid also contained chloramphenicol acetyltransferase for resistance during plating (CmR).
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[[File:AmilCP in pSB1C3 Map.png|200px|thumb|centre|pSB1C3 map]]
  
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===Applications of BBa_K2078002===
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====Observations and Inferences====
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Our E.coli grew slower than other transformants and we were also unable to recover as much plasmid by miniprep. The use of this strong consensus promoter slows down expression times of the AmilCP [[Part:BBa_K592025]] in E-Coli by around 24hours. This image shows a pellet from 5ml LB broth with transformed E.coli and confirms that the part is expressed, but the amount of pellet was much less than we would normally expect to get from 5ml.
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[[File:Pellet from 5ml LB broth with transformed Ecoli.jpeg|200px|thumb|centre|Pellet from 5ml LB broth]]
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The delay in E-Coli growth is also shown by this 24hour streak plate. The chromoprotein is only just becoming visible suggesting that such a strong consensus sequence slows down both the expression time of the part and growth time of E-Coli.
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[[File:Ecoli streak plate after 24 hours.jpeg|200px|thumb|centre|Transformed E-Coli streak plate after 24hours]].
  
 
===User Reviews===
 
===User Reviews===

Revision as of 13:41, 9 October 2016

Applications of BBa_K2078002

Experience of CLSB-UK-2016 iGEM team with Part:BBa_K2078002

Experimental Design

This consensus promoter was assembled with the reporter protein, blue-purple chromoprotein AmilCP. The construct was carried on the pSB1C3 plasmid for expression and amplification in E-Coli. The plasmid also contained chloramphenicol acetyltransferase for resistance during plating (CmR).

pSB1C3 map


Observations and Inferences

Our E.coli grew slower than other transformants and we were also unable to recover as much plasmid by miniprep. The use of this strong consensus promoter slows down expression times of the AmilCP Part:BBa_K592025 in E-Coli by around 24hours. This image shows a pellet from 5ml LB broth with transformed E.coli and confirms that the part is expressed, but the amount of pellet was much less than we would normally expect to get from 5ml.

Pellet from 5ml LB broth

The delay in E-Coli growth is also shown by this 24hour streak plate. The chromoprotein is only just becoming visible suggesting that such a strong consensus sequence slows down both the expression time of the part and growth time of E-Coli.

Transformed E-Coli streak plate after 24hours
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User Reviews

UNIQ9e218af94dfb0e6e-partinfo-00000000-QINU UNIQ9e218af94dfb0e6e-partinfo-00000001-QINU