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US 5,270,181 AGranted
Peptide and protein fusions to thioredoxin and thioredoxin-like molecules
AssigneeGenetics Institute, Inc.
InventorsJohn McCoy, Edward R. LaVallie
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AssigneeGenetics Institute, Inc.
InventorsJohn McCoy, Edward R. LaVallie
Estimating the term…
This invention provides a fusion molecule comprising a DNA sequence encoding a thioredoxin-like protein fused to the DNA sequence encoding a selected heterologous peptide or protein. The peptide or protein may be fused to the amino terminus of the thioredoxin-like molecule, the carboxyl terminus of the thioredoxin-like molecule, or within the thioredoxin-like molecule, for example at the active-site loop of said molecule. Expression of this fusion molecule under the control of a regulatory sequence capable of directing its expression in a desired host cell, produces high levels of stable and soluble fusion protein. The fusion protein, located in the bacterial cytoplasm, may be selectively released from the cell by osmotic shock or freeze/thaw procedures. It may be optionally cleaved to liberate the soluble, correctly folded heterologous protein from the thioredoxin-like portion.
Fusion DNA having a formula selected from the group consisting of R1-R2, R2-R1, R1-L-R2 and R2-L-R1 wherein, R1 is thioredoxin-like DNA;
Fusion DNA comprising (1) thioredoxin-like DNA, said thioredoxin-like DNA characterized by encoding a polypeptide that (a) has a three-dimensional structure substantially similar to that of E. coli thioredoxin and (b) contains an active site loop functionally and structurally equivalent to the double cysteine containing active site loop of E. coli thioredoxin;
Fusion DNA having a formula selected from the group consisting of R1-R2, R2-R1, R1-L-R2 and R2-L-R1 wherein, R1 is DNA encoding E. coli thioredoxin;
Fusion DNA having a formula R1-L-R2, wherein R1 is DNA encoding E. coli thioredoxin;
Dependencies are inferred from the claim text.