Mutant proteinase with reduced self-cleavage activity and method of purification
AssigneeYale University
InventorsJennifer A. Doudna, Louise J. Lucast, Robert T. Batey
Life of this patent
Estimating the term…
Abstract
The present invention provides a mutant 27 kDa NIa proteinase having reduce self-cleavage activity relative to the self-cleavage activity of its wild-type proteinase. The mutant the same substrate cleavage activity as the wild-type proteinase but is more stable than the wild-type proteinase. The present invention also provides a method of obtaining large quantities of active 27 kDa NIa proteinase for use as a tool for purification of other proteins.
Claims23
An isolated active mutant tobacco etch virus (TEV) proteinase having a molecular weight of about 27 kDa and reduced self-cleavage activity relative to the self-cleavage activity of wild-type TEV proteinase.
An isolated nucleic acid molecule comprising a sequence encoding an active mutant TEV proteinase having a molecular weight of about 27 kDa and reduced self-cleavage activity relative to the self-cleavage activity of wild-type TEV proteinase.
A nucleic acid encoding a fusion protein comprising an active mutant TEV proteinase, having a molecular weight of about 27 kDa and reduced self-cleavage activity relative to the self-cleavage activity of wild-type TEV proteinase, fused to a heterologous protein.
An isolated active mutant tobacco etch virus (TEV) proteinase comprising an amino acid sequence in which the residue corresponding to Ser 219 of the wild-type TEV proteinase is replaced with another residue and wherein the isolated active mutant TEV proteinase has a reduced self-cleavage activity relative to the wild-type TEV proteinase.
Dependencies are inferred from the claim text.