Difference between revisions of "Part:BBa K4041009"

 
 
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This is for the surface display of the protein Yafl-Ulp1 to cleave the SUMO-protein partner into SUMO and protein. Which in turn purifies the protein of choice without the use of tags. This method allows easy centrifugation to purify protein in a relatively pure output.
 
This is for the surface display of the protein Yafl-Ulp1 to cleave the SUMO-protein partner into SUMO and protein. Which in turn purifies the protein of choice without the use of tags. This method allows easy centrifugation to purify protein in a relatively pure output.
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<br><br>
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https://static.igem.org/mediawiki/parts/8/80/HKIS--cleav.png<br>
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<br><br>Figure 2, Illustration of Cleavage system (brown Yafl Ulp1, Red fusion protein)<br><br>
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Source: https://amb-express.springeropen.com/articles/10.1186/s13568-020-00999-4
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https://amb-express.springeropen.com/articles/10.1186/s13568-020-00999-4
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This system does not work well with system that does cannot bear the toxicity, like the BL21 (DE3) -gold cells, which has a high mRNA stability. <br><br>
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<br>
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This system does not work well with system that does cannot bear the toxicity, like the BL21 (DE3) -gold cells, which has a high mRNA stability.  
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https://static.igem.org/mediawiki/parts/b/bb/HKIS--yaflexpression.png<br><br>
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Figure 2, showing the expression of Yafl from our own test, pET-LO-SUMO and pET-LO-SUMO-tachyplesin system.
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https://static.igem.org/mediawiki/parts/b/bb/HKIS--yaflexpression.png
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<br> <br> <br> From the picture, it is suggested to use lysis resistant strains and other strains with high toxicity tolerance for the IPTG induction of the peptide, preferably in high enrichment broth including terrific broth or magic media.
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Figure 1, showing the expression of Yafl, pET-LO-SUMO and pET-LO-SUMO-tachyplesin system.
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From the picture, it is suggested to use lysis resistant strains and other strains with high toxicity tolerance for the IPTG induction of the peptide, preferably in high enrichment broth including terrific broth or magic media.
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Latest revision as of 20:49, 21 October 2021


pET-Yafl-Ulp1

This is for the surface display of the protein Yafl-Ulp1 to cleave the SUMO-protein partner into SUMO and protein. Which in turn purifies the protein of choice without the use of tags. This method allows easy centrifugation to purify protein in a relatively pure output.

HKIS--cleav.png


Figure 2, Illustration of Cleavage system (brown Yafl Ulp1, Red fusion protein)

Source: https://amb-express.springeropen.com/articles/10.1186/s13568-020-00999-4

This system does not work well with system that does cannot bear the toxicity, like the BL21 (DE3) -gold cells, which has a high mRNA stability.


HKIS--yaflexpression.png

Figure 2, showing the expression of Yafl from our own test, pET-LO-SUMO and pET-LO-SUMO-tachyplesin system.


From the picture, it is suggested to use lysis resistant strains and other strains with high toxicity tolerance for the IPTG induction of the peptide, preferably in high enrichment broth including terrific broth or magic media.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 2185
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 410
    Illegal BamHI site found at 1053
    Illegal BamHI site found at 1203
    Illegal BamHI site found at 1215
    Illegal BamHI site found at 1233
    Illegal BamHI site found at 1239
    Illegal XhoI site found at 2194
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 883
    Illegal AgeI site found at 1000
    Illegal AgeI site found at 2068
  • 1000
    COMPATIBLE WITH RFC[1000]