Part:BBa_K4041016
Expression Cassette for 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
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 1189
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 3445
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1147
Illegal BglII site found at 1670
Illegal BamHI site found at 2313
Illegal BamHI site found at 2463
Illegal BamHI site found at 2475
Illegal BamHI site found at 2493
Illegal BamHI site found at 2499
Illegal XhoI site found at 3454 - 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 1189
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 1189
Illegal AgeI site found at 2143
Illegal AgeI site found at 2260
Illegal AgeI site found at 3328 - 1000COMPATIBLE WITH RFC[1000]
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