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AssigneeQueen's University at Kingston
InventorsLaurie A. Graham, Yih-Cherng Liou, Virginia K. Walker, Peter L. Davies
Estimating the term…
A novel class of thermal hysteresis (antifreeze) proteins (THP) that have up to 100 times the specific activity of fish antifreeze proteins has been isolated and purified from the mealworm beetle, Tenebrio molitor. Internal sequencing of the proteins, leading to cDNA cloning and production of the protein in bacteria has confirmed the identity and activity of the 8.4 to 10.7 kDa THP. They are novel Thr- and Cys-rich proteins composed largely of 12-amino-acid repeats of cys-thr-xaa-ser-xaa-xaa-cys-xaa-xaa-ala-xaa-thr. At a concentration of 55 μg/mL, the THP depressed the freezing point 1.6° C. below the melting point, and at a concentration of ˜1 mg/mL the THP or its variants can account for the 5.5° C. of thermal hysteresis found in Tenebrio larvae. The THP function by an adsorption-inhibition mechanism and produce oval-shaped ice crystals with curved prism faces.
An isolated nucleic acid encoding an antifreeze protein, said protein defined as follows:
An isolated nucleic acid which specifically hybridizes to SEQ ID NO:2 under stringent conditions.
An isolated nucleic acid encoding at least 1 repeat of a 12 contiguous amino acid motif from a purified Tenebrio molitor antifreeze protein which specifically binds to an antibody directed against antifreeze proteins selected from the group consisting of YL-1, YL-2 YL-3 and YL-4.
An isolated antifreeze protein, said protein:
An antibody, specifically immunoreactive under Immunologically reactive conditions, to an antifreeze protein, said protein having at least 1 repeat of a sequence comprising the motif of SEQ ID NO:1.
An organism into which, or into an ancestor of which, an exogenous nucleic acid sequence which specifically hybridizes under stringent conditions to SEQ ID NO:4 has been introduced and the organism translates the exogenous nucleic acid into an antifreeze protein.
The organism of claim 18, wherein said fish is a member of the family Salmonidae.
A method for decreasing the freezing point of an aqueous solution, said method comprising the addition of an antifreeze protein having greater than 1 repeat of the contiguous amino acid motif of SEQ ID NO:1, to said aqueous solution.
Dependencies are inferred from the claim text.