Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Shuyan Tang
Date created: 2015-09-05 11:00:00
Date modified: 2015-09-18 09:27:28
LIP2 prepro + E. coli ribosomal protein L2 (203-273aa) + YLcwp3 Fusion
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K1592009_sequence (Version 1) |
Description
This is the cell display system of Yarrowia lipolytica, composed of LIP2 prepro, interest protein, and YLcwp3.LIP prepro is signal peptide used to secrete the interest protein out of the cell, and the YLcwp3 is the anchor domain binding the interest protein to the cell wall of yeast.We use this system to display silica-tag and test its binding characteristics.
E.coli ribosomal protein L2 was found to bind tightly to silicon particles, which have surfaces that are oxidized to silica. This L2 silica-binding tag, called the 'Si-tag', can be used for one-step targeting of functional proteins on silica surfaces. The silica-binding domains of E. coli L2 was mapped to amino acids 1???60, 61-202 and 203???273, called Si-tag1, Si-tag2, Si-tag3. We respectively test the silica-binding characteristics of this three regions and their combinations.
Notes
In order to replace the domains more conveniently, we respectivey added BamHI, SalI, NdeI between LIP2 prepro, E. coli ribosomal protein L2, GS linker and YLcwp3.Source
E. coli ribosomal protein L2 was synthesized by IDT, LIP2 prepro and YLcwp3 were cloning from the plasmid JMP62, and we fused this three sequences.| Sequence Annotation | Location | Component / Role(s) |
| LIP2 prepro BamHI His6 tag Si-tag 3 SalI GS Linker YLcwp3 NdeI | 1,99 100,105 106,123 124,339 340,345 346,372 379,762 373,378 | CDS feature/protein sequence_feature feature/misc tag feature/tag CDS feature/protein feature/misc sequence_feature tag feature/tag feature/protein CDS sequence_feature feature/misc |