Part:BBa_K4041010

pET-LO-SUMO This is a incomplete part without desired protein fused to SUMO partner, the required protein need to be cloned into the NdeI via NEBuilder. This is for the surface display of the protein SUMO to be cleaved by the Yafl-Ulp1-protein partner into SUMO and protein into the cleavage solution outside of the cell. Which in turn purifies the protein of choice without the use of tags but only centrifugation. 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

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 560
  Illegal XhoI site found at 766
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 370
  
• 1000
  COMPATIBLE WITH RFC[1000]