Team:HUST-China/Part Collection
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Best Part Collection:
This year we had 26 parts documented and already sent them to the registry of Standard Biological Parts. Particularly,we did well in the standardization of E.coli 50S ribosomal protein (Si-tag) with the different combination of its domains:1,2,3,1+2,1+3,2+3,1+2+3,1L3.
The protein has been found from several bacterial strains and proved to have a silica-binding property. To make it a more thouroghly functional part,we added a signal peptide LIP2 prepro and cell wall protein of Yarrowia.Lipolytica YLcwp3 respectively at the N end and C end of the Si-tags.
The surface display strategy expressed in eukaryotic cells can immobilize the silica affinitive protein on the surface of the cell wall. We achived a considerable characterazation results with fluorescence immunoassay to test the surface display system and glass binding test to verify the proteins’s affinity to silica solid. The former one showed that we succeeded in displaying the silica-tag onto cell surface and the latter proved each protein domain had different binding ability. So we can choose different combinations of Si-tag domains to satisfy different requirement of binding intensity thus making a more flexible application.
Here is the collection list (click Part Number to see more details)
| Part Number | Description | Abbreviation |
|---|---|---|
| BBa_K1592007 | LIP prepro + E. coli ribosomal protein L2 (1-60) + YLcwp3 Fusion | Si-tag1-his |
| BBa_K1592008 | LIP prepro + E. coli ribosomal protein L2 (61-202) + YLcwp3 Fusion | Si-tag2-his |
| BBa_K1592009 | LIP prepro + E. coli ribosomal protein L2 (203-273) + YLcwp3 Fusion | Si-tag3-his |
| BBa_K1592010 | LIP2 prepro + E. coli ribosomal protein L2 (1-202)+ YLcwp3 Fusion | ST12 |
| BBa_K1592011 | LIP prepro + E. coli ribosomal protein L2 (61-273) + YLcwp3 Fusion | ST23 |
| BBa_K1592012 | LIP prepro + E. coli ribosomal protein L2 (1-60,203-273) + YLcwp3 Fusion | ST13 |
| BBa_K1592013 | LIP prepro + E. coli ribosomal protein L2 (1-60,GS linker,202-273) + YLcwp3 | ST1L3-his |
| BBa_K1592014 | LIP prepro + E. coli ribosomal protein L2 (1-273) + YLcwp3 | ST123 |