Part:BBa_K4041009
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.
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.
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 2185
- 21INCOMPATIBLE 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 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 883
Illegal AgeI site found at 1000
Illegal AgeI site found at 2068 - 1000COMPATIBLE WITH RFC[1000]
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