Difference between revisions of "Part:BBa K2333005"

Latest revision as of 18:08, 29 October 2017

mf-Lon Protein Degradation Tag E (medium-weak)

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 the second weakest, and has the second slowest 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.

These parts are available in the William and Mary UNS format of: Tag-B0015(Double Terminator)-DoubleStop Codon which enables easy cloning using Gibson, Golden Gate or 3A assembly. See part series K2333401-K2333406 or BBa_K2333405 for this degradation tag.

Note: pdt E varies from pdt A in that residues 13-15 change from “PTF” to “APN”.

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]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2333005 short</partinfo>
-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 the second weakest, and has the second slowest 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.
-Note: pdt #3d varies from pdt #3 in that residues 13-15 change from “PTF” to “APN”.
+Of this series of tags, this tag is the second weakest, and has the second slowest 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.
+These parts are available in the William and Mary UNS format of: Tag-B0015(Double Terminator)-DoubleStop Codon which enables easy cloning using Gibson, Golden Gate or 3A assembly. See part series K2333401-K2333406 or <partinfo>K2333405</partinfo> for this degradation tag.
+Note: pdt E varies from pdt A in that residues 13-15 change from “PTF” to “APN”.
 <!-- Add more about the biology of this part here