Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Luis Mario Leal Garza
Date created: 2012-09-25 11:00:00
Date modified: 2015-05-08 01:13:48
Secretion Sequence+antiIgE Scfv+GFP+Etag
| Types | DnaRegion |
| Roles | CDS Coding |
| Sequences | BBa_K942005_sequence (Version 1) |
Description
This part uses a secretion signal for yeasts. Also the main section of this part is a fusion protein made from a murine scFv specific to human IgE at the N-terminal and a yeast-enhanced green fluorescent protein (yEGFP) at the C-terminal part of the protein. (Christian Lupinek, 2009)This fusion protein weights about 53kDa. It binds to human IgE and works as a GFP also. The C-terminal Etag was not used in our project, instead a 30kDa ultrafiltration was done. The scFv part of the fusion works by binding human IgE. (Gasteiger E., 2005)
About Safety
The part itself does not present any biohazard characteristic, but it is important to remember that it is designed to work in Pichia pastoris and that a biosecurity chamber is recommended to handle this organism. Its secretion to the media should not suppose any problem.
Notes
This part is codon-optimized to be expressed in yeast. This part has a secretion signal at is N-terminal and an affinity purification tag for easy recovery. The fusion between scFv IgE and GFP is achieved by a (GGGGS)4 linker.Source
The main section of this part is a fusion protein made from a murine scFv specific to human IgE at the N-terminal and a yeast-enhanced green fluorescent protein (yEGFP) at the C-terminal part of the protein. (Christian Lupinek, 2009)Christian Lupinek et al. (2009) Trimolecular Complex Formation of IgE, FcERI, and a Recombinant Nonanaphylactic Single-Chain Antibody Fragment with High Affinity for IgE. The Journal of Immunology 4817-4829