Difference between revisions of "Part:BBa K2333004"
Line 4: Line 4:
  
 
This protein degradation tag (pdt) is degraded by the Lon protease from Mycoplasma florum (mf-Lon). <b>LINK TO mf-Lon goes here </b>The Lon protease efficiently recognize and degrade C-terminal ssrA-tagged proteins, functioning in a similar way as ClpXP protease in most gram-negative bacteria. This Lon protease system is orthogonal to E. coli’s endogenous protein degradation machinery. This tag is part of a series of 6 tags (labeled #3 and #3a-#3e). pdt#3 is used as the parental tag to create #3a-#3e by changing the pdt residues 13–15 for mutagenesis because this region is essential for Lon-mediated degradation. Of this series of tags, this tag is relatively weak, and has a slow degradation rate. BBa_K2333001-Bba_K2333006 comprise this tag series. See Collins et al. 2014 "Tunable Protein Degradation in Bacteria" for the original design via mutagenesis, and for their characterization of degradation rates.
 
This protein degradation tag (pdt) is degraded by the Lon protease from Mycoplasma florum (mf-Lon). <b>LINK TO mf-Lon goes here </b>The Lon protease efficiently recognize and degrade C-terminal ssrA-tagged proteins, functioning in a similar way as ClpXP protease in most gram-negative bacteria. This Lon protease system is orthogonal to E. coli’s endogenous protein degradation machinery. This tag is part of a series of 6 tags (labeled #3 and #3a-#3e). pdt#3 is used as the parental tag to create #3a-#3e by changing the pdt residues 13–15 for mutagenesis because this region is essential for Lon-mediated degradation. Of this series of tags, this tag is relatively weak, and has a slow degradation rate. BBa_K2333001-Bba_K2333006 comprise this tag series. See Collins et al. 2014 "Tunable Protein Degradation in Bacteria" for the original design via mutagenesis, and for their characterization of degradation rates.
 +
 
Note: pdt #3a varies from pdt #3 in that residues 13-15 change from “PTF” to “AQP”.
 
Note: pdt #3a varies from pdt #3 in that residues 13-15 change from “PTF” to “AQP”.
  

Revision as of 22:48, 14 June 2017


mf-Lon Protein Degradation Tag D (medium)

This protein degradation tag (pdt) is degraded by the Lon protease from Mycoplasma florum (mf-Lon). LINK TO mf-Lon goes here The Lon protease efficiently recognize and degrade C-terminal ssrA-tagged proteins, functioning in a similar way as ClpXP protease in most gram-negative bacteria. This Lon protease system is orthogonal to E. coli’s endogenous protein degradation machinery. This tag is part of a series of 6 tags (labeled #3 and #3a-#3e). pdt#3 is used as the parental tag to create #3a-#3e by changing the pdt residues 13–15 for mutagenesis because this region is essential for Lon-mediated degradation. Of this series of tags, this tag is relatively weak, and has a slow degradation rate. BBa_K2333001-Bba_K2333006 comprise this tag series. See Collins et al. 2014 "Tunable Protein Degradation in Bacteria" for the original design via mutagenesis, and for their characterization of degradation rates.

Note: pdt #3a varies from pdt #3 in that residues 13-15 change from “PTF” to “AQP”.

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]