Difference between revisions of "Part:BBa K2333003"

Revision as of 15:15, 29 October 2017

mf-Lon Protein Degradation Tag C (medium-strong)

This protein degradation tag (pdt) is degraded by the Lon protease from Mycoplasma florum (mf-Lon) (BBa_K2333011).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 A thru F). pdt A is used as the parental tag to create tags B-F 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 moderately strong, and has high 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 B varies from pdt A in that residues 13-15 change from “PTF” to “RAI”.

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]

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 <partinfo>BBa_K2333003 short</partinfo>
-This protein degradation tag (pdt) is degraded by the Lon protease from Mycoplasma florum (mf-Lon) (<partinfo>Bba_K2333011</partinfo>).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 A thru F). pdtA is used as the parental tag to create tags B-F 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 moderately strong, and has high 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) (<partinfo>Bba_K2333011</partinfo>).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 A thru F). pdt A is used as the parental tag to create tags B-F 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 moderately strong, and has high 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 B varies from pdt A in that residues 13-15 change from “PTF” to “RAI”.