Difference between revisions of "Part:BBa K4241024"

 
 
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<partinfo>BBa_K4241024 short</partinfo>
 
<partinfo>BBa_K4241024 short</partinfo>
  
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This is the expression of a T7 tagged SUMO-CeB fusion, which the RFP could be his-tag pulled down and purified via Ulp1 cleavage and second round of His-tag pulldown.
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<partinfo>BBa_K4241024 SequenceAndFeatures</partinfo>
  
  
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<!-- Add more about the biology of this part here
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===Overview===
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===Usage and Biology===
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This composite part is for the expression of a T7 tagged, SUMO-CeB fusion protein. This allows for the production and purification of the Cecropin B via a double His-tag purification scheme by cleaving SUMO with Ulp1.
  
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<!-- -->
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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This is a bacterial expression system for SUMO-RFP fusion protein. By expressing the T7 tagged SUMO-RFP fusion protein via an enhanced T7pCONS system, we are able to increase the expression yield of the protein. The nature of the SUMO-tag further allows for a simplified purification scheme.
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<partinfo>BBa_K4241024 SequenceAndFeatures</partinfo>
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<br/>
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Purification scheme:<br/>
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1. Express the fusion protein in E. coli and sonicate cells <br/>
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2. His-pulldown the cell lysate <br/>
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3. Cleave with Ulp1 <br/>
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4. His-pulldown to seperate uncleaved proteins and non-target proteins.
  
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===Parts that comprise this composite component===
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Enhance-T7pro-RBS-Start-6xHis-T7tag - Part:BBa_K4241016 <br/>
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SUMO (Yeast with mutation) - Part:BBa_K4241022 <br/>
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Cecropin B bacterial expression - Part:BBa_K4241020 <br/>
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Double stop codon - Part:BBa_K4241017 <br/>
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double terminator (B0010-B0012) - Part:BBa_B0015 <br/>
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 16:25, 9 October 2022


6xHis_SUMO_CeB


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 155
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 301
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Overview

This composite part is for the expression of a T7 tagged, SUMO-CeB fusion protein. This allows for the production and purification of the Cecropin B via a double His-tag purification scheme by cleaving SUMO with Ulp1.

Usage and Biology

This is a bacterial expression system for SUMO-RFP fusion protein. By expressing the T7 tagged SUMO-RFP fusion protein via an enhanced T7pCONS system, we are able to increase the expression yield of the protein. The nature of the SUMO-tag further allows for a simplified purification scheme.
Purification scheme:
1. Express the fusion protein in E. coli and sonicate cells
2. His-pulldown the cell lysate
3. Cleave with Ulp1
4. His-pulldown to seperate uncleaved proteins and non-target proteins.

Parts that comprise this composite component

Enhance-T7pro-RBS-Start-6xHis-T7tag - Part:BBa_K4241016
SUMO (Yeast with mutation) - Part:BBa_K4241022
Cecropin B bacterial expression - Part:BBa_K4241020
Double stop codon - Part:BBa_K4241017
double terminator (B0010-B0012) - Part:BBa_B0015