Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Shuyan Tang
Date created: 2015-09-04 11:00:00
Date modified: 2015-09-18 09:26:29
LIP prepro + E. coli ribosomal protein L2 (1-60aa) + YLcwp3 Fusion
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K1592007_sequence (Version 1) |
Description
This is the cell display fusion protein 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.
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. We respectively test the silica-binding characteristics of this three regions and their combinations. E. coli ribosomal protein L2 (1-60) is the amino acids 1???60 of E. coli ribosomal protein L2, we call it si-tag1.
Notes
In order to replace more convenient, we respectively added BamHI, SalI, NdeI between LIP2 prepro, E. coli ribosomal protein L2 (1-60), GS linker, YLcwp3.Source
E. coli ribosomal protein L2 was synthesized by IDT.LIP2 prepro and YLcwp3 were cloning from the plasmid JMP62 of our lab.
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K1592000 BamHI Si-tag1 SalI GS linker NdeI YLcwp3 BBa_K1592002 LIP2 prepro his-tag | 1,99 100,105 124,303 304,309 310,336 337,342 343,708 343,708 1,99 106,123 | engineered_region feature/BioBrick sequence_feature feature/misc CDS feature/protein feature/misc sequence_feature tag feature/tag sequence_feature feature/misc feature/protein CDS feature/BioBrick engineered_region feature/protein CDS tag feature/tag |