Abstract 2022 Cloning a codon-optimized version of the Rhagnium Inquisitor Antifreeze Protein gene into the pTYB12 expression vector

The purpose of this project is to clone and express a synthetic version of antifreeze protein, RiAFP, from the arctic beetle, Rhagnium Inquisitor. The coding sequence for the RiAFP was codon-optimized for expression in E. Coli. Natively expressed RiAFP prevents the arctic beetle's body fluids from freezing by binding to the surface of ice crystals and inhibits the ability for the crystals to grow and expand.1 This protein exhibits a unique beta fold. The Beta sheets stack on top of each other in a repeating and parallel form.