Difference between revisions of "Part:BBa K1075021"
 
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<partinfo>BBa_K1075021 short</partinfo>
 
<partinfo>BBa_K1075021 short</partinfo>
  
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ecssrA(DAS+4)-TT
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The part contains the mutated E. coli(ec)ssrA (DAS+4) tag and a double terminator.
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===construction===
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The part contains the mutated ssrA (DAS+4) tag and a double terminator.
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===biology===
 
===biology===
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The ssrA tag is a short peptide sequence, which is fused to the C-terminus of proteins, which should be degraded. It relates to the protease ClpXP complex in E.coli and it also allows the adaptor proteins SspB binding and delivering substrates to the proteases in order to make the process more efficient. The mutated ssrA (DAS+4) weakens a direct binding between proteases and ssrA and increases the dependance of sspB.
 
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The double terminator stops the transcription at this point.
 
  
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The (ec)ssrA tag is a short peptide sequence that can be fused to the C-terminus of proteins which should be degraded. The adaptor protein SspB binds to it and delivers the substrates to the protease ClpXP complex in E. coli. The mutation (DAS+4) in (ec)ssrA prevents the degradation of the ssrA-tagged protein without the presence of sspB and therefore makes the system more specific and controllable. [http://www.ncbi.nlm.nih.gov/pubmed/16762842]
  
 
===application===
 
===application===
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As we want to control protein degradation by controlling the function of SspB, we tag the red fluorescent protein mCherry with ssrA (DAS+4) to measure the degradation rate.
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The (ec)ssrA(DAS+4) tag can be fused to proteins so that they will be delivered to the ClpXP protease and degraded.  
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Latest revision as of 01:12, 5 October 2013

E. coli ssrA(DAS+4)-TT

The part contains the mutated E. coli(ec)ssrA (DAS+4) tag and a double terminator.

biology

The (ec)ssrA tag is a short peptide sequence that can be fused to the C-terminus of proteins which should be degraded. The adaptor protein SspB binds to it and delivers the substrates to the protease ClpXP complex in E. coli. The mutation (DAS+4) in (ec)ssrA prevents the degradation of the ssrA-tagged protein without the presence of sspB and therefore makes the system more specific and controllable. [http://www.ncbi.nlm.nih.gov/pubmed/16762842]

application

The (ec)ssrA(DAS+4) tag can be fused to proteins so that they will be delivered to the ClpXP protease and degraded.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 40
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]