Difference between revisions of "Part:BBa K174002"

 
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<partinfo>BBa_K174002 short</partinfo>
 
<partinfo>BBa_K174002 short</partinfo>
  
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The purpose of this system is to control the expression of sspB, the degradation controller. The system is inducible by arabinose, therefore we can alter the arabinose levels to regulate the degradation of proteins with sspB degradation tag
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The purpose of this parts is to control the expression of sspB, the degradation controller.  
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Mutations on ssrA degradation tags weakens the recognition of the tagged protein by ClpX. Modified versions require sspB adaptor protein for the degradation. Hence when sspB protein is expressed, the proteins tagged with modified version of ssrA tag are targeted for degradation, otherwise they remain stable (1).
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This device is inducible by arabinose, therefore we can alter the arabinose levels to regulate the degradation of proteins tagged with the modified version of ssrA degradation tags.
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Wild type E. coli ssrA tag is AANDENY-ALAA (SspB recognition site – ClpX recognition site).
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As an example of the tag, we constructed [[Part:BBa_K174003]] with Hin tagged with the modified version of ssrA tag below.
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AANDENY-SENY-ALGG (SspB recognition site – SENY +4 Linker - ClpX recognition site)
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HJowever it should also be noted that different degradation tags may have effect on the activity of different proteins (1).
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For more information about this part, go to Newcastle iGEM2009 [http://2009.igem.org/Team:Newcastle/Stochasticity Stochasticity] and [http://2009.igem.org/Team:Newcastle/Modelling Modelling] pages.
  
 
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<partinfo>BBa_K174002 parameters</partinfo>
 
<partinfo>BBa_K174002 parameters</partinfo>
 
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===References===
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#Griffith, K. L., and A. D. Grossman. 2008. Inducible protein degradation in Bacillus subtilis using heterologous peptide tags and adaptor proteins to target substrates to the protease ClpXP. Mol. Microbiol. 70:1012-1025.

Revision as of 20:50, 19 October 2009

Arabinose controlled protein degradation device

The purpose of this parts is to control the expression of sspB, the degradation controller.

Mutations on ssrA degradation tags weakens the recognition of the tagged protein by ClpX. Modified versions require sspB adaptor protein for the degradation. Hence when sspB protein is expressed, the proteins tagged with modified version of ssrA tag are targeted for degradation, otherwise they remain stable (1).

This device is inducible by arabinose, therefore we can alter the arabinose levels to regulate the degradation of proteins tagged with the modified version of ssrA degradation tags.


Wild type E. coli ssrA tag is AANDENY-ALAA (SspB recognition site – ClpX recognition site).

As an example of the tag, we constructed Part:BBa_K174003 with Hin tagged with the modified version of ssrA tag below.

AANDENY-SENY-ALGG (SspB recognition site – SENY +4 Linker - ClpX recognition site)

HJowever it should also be noted that different degradation tags may have effect on the activity of different proteins (1).

For more information about this part, go to Newcastle iGEM2009 [http://2009.igem.org/Team:Newcastle/Stochasticity Stochasticity] and [http://2009.igem.org/Team:Newcastle/Modelling Modelling] pages.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. Griffith, K. L., and A. D. Grossman. 2008. Inducible protein degradation in Bacillus subtilis using heterologous peptide tags and adaptor proteins to target substrates to the protease ClpXP. Mol. Microbiol. 70:1012-1025.