Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Shuyan Tang
Date created: 2015-09-07 11:00:00
Date modified: 2015-09-18 09:36:35
LIP prepro + E. coli ribosomal protein L2 (1-60,GS linker,202-273aa) + YLcwp3
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K1592013_sequence (Version 1) |
Description
__NOTOC__This is the cell display system of Yarrowia lipolytica, composed of LIP 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.
Here we use this system to display Silica-tag on cell wall of Yarrowia lipolytica to binding the silica.
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 and 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 YLcwp3Source
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 1L3 SalI GS Linker NdeI YLcwp3 GS Linker | 1,99 100,105 106,123 124,534 535,540 541,585 586,591 592,957 304,318 | CDS feature/protein feature/misc sequence_feature feature/tag tag feature/protein CDS feature/misc sequence_feature feature/tag tag sequence_feature feature/misc CDS feature/protein tag feature/tag |