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
proU+RiAFP+Term
| Types | DnaRegion |
| Roles | Generator engineered_region |
| Sequences | BBa_K942012_sequence (Version 1) |
Description
This protein has an approximate weight of 13.7kDa and is better visualized in a SDS-PAGE with Tris-Tricine gel buffer, because of its low molecular weight. It can be purified by affinity with its 6xHis tag. An induction with 300mM of NaCl presents good expression. (Gasteiger E., 2005)About Safety
The RiAFP has no reported toxicity or hazardousness, but its important to remember to have the basic biosafety precautions using a biosecurity chamber when handling any transformed strains. The inductor also presents no hazard as it is just common salt.
Notes
The RiAFP is expressed when around 300mM NaCl added. This part was designed by adding a 6xHis tag for affinity purification to the anti freeze protein from Raghium inquisitor. This part was codon optimized for its expression in E.coli.Source
The Raghium inquisitor Anti Freeze Protein (RiAFP) is a 137 AA protein that prevents the growth of ice crystals. The proU promoter is from E.coli and works activating transcription when the bacterium is osmotically stressed. (Kristiansen E, 2011)Kristiansen, E. et all (2011) Structural characteristics of a novel antifreeze protein from the longhorn beetle Rhagium inquisitor. Insect Biochem Mol Biol. 41(2):109-17.