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
pBAD+OmpA+RiAFP+Term
| Types | DnaRegion |
| Roles | engineered_region Generator |
| Sequences | BBa_K942013_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. (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 presents no hazard as is arabinose.
Notes
The RiAFP is expressed when around 0.1% L-Arabinose is added. This part was designed by adding a 6xHis tag for affinity purification to the anti freeze protein from Raghium inquisitor and the signal sequence of the ompA gene for periplasmic secretion. 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. This protein is produced natively in the beetle Raghium inquisitor. This part includes the OmpA signal sequence native from E.coli and the pBAD L-arabinose inducible promoter. (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.