Difference between revisions of "Part:BBa K1051206"
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<h3>Purpose</h3>
 
<h3>Purpose</h3>
−
SsrA degradation tag in E.coli, add TAATAA to C-terminal of Biobrick M0050.
+
SsrA degradation tag in <i />E. coli, add TAATAA to C-terminal of Biobrick M0050.
 
<br />
 
<br />
 
<h3>Principle</h3>
 
<h3>Principle</h3>
−
'''SsrA degradation tag'''. In E.coli, the adaptor SspB tethers ssrAtagged substrates to the ClpXP protease, causing a modest increase in their rate of degradation. Which means, a variation of the WT SsrA tag sequence will accelerate the degradation of proteins when fused to their C-terminal. Thus the degradation rates are dependent on concentration of proteases and binding mediators. In order to fuse degradation tags freely on the C-terminal of protein, we add TAATAA to the tail of M0050 to construnt this part.<br />
+
'''SsrA degradation tag'''. In E. coli, the adaptor SspB tethers ssrAtagged substrates to the ClpXP protease, causing a modest increase in their rate of degradation. Which means, a variation of the WT SsrA tag sequence will accelerate the degradation of proteins when fused to their C-terminal. Thus the degradation rates are dependent on concentration of proteases and binding mediators. In order to fuse degradation tags freely on the C-terminal of protein, we add TAATAA to the tail of M0050 to construnt this part.<br />
 
We constructed the measurement pathway of tag K1051257(contains J04500, K1051000, K1051206) to test the rates of degradation of tagged proteins. J04450 was used as positive control because of the same promoter and fluorescent protein.  
 
We constructed the measurement pathway of tag K1051257(contains J04500, K1051000, K1051206) to test the rates of degradation of tagged proteins. J04450 was used as positive control because of the same promoter and fluorescent protein.  
 
<br />
 
<br />
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<h3>References</h3>
 
<h3>References</h3>
−
(1)McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701.
+
[1]McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701.</h3>
−
(2)Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240
+
[2]Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240

Revision as of 16:10, 27 September 2013

The degradation tag in E. coil ,M0050 with TAATAA.

Purpose

SsrA degradation tag in <i />E. coli, add TAATAA to C-terminal of Biobrick M0050.

Principle

SsrA degradation tag. In E. coli, the adaptor SspB tethers ssrAtagged substrates to the ClpXP protease, causing a modest increase in their rate of degradation. Which means, a variation of the WT SsrA tag sequence will accelerate the degradation of proteins when fused to their C-terminal. Thus the degradation rates are dependent on concentration of proteases and binding mediators. In order to fuse degradation tags freely on the C-terminal of protein, we add TAATAA to the tail of M0050 to construnt this part.
We constructed the measurement pathway of tag K1051257(contains J04500, K1051000, K1051206) to test the rates of degradation of tagged proteins. J04450 was used as positive control because of the same promoter and fluorescent protein.

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]McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701.</h3> [2]Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240