Peptide and protein fusions to thioredoxin, thioredoxin-like mol

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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4351723, 435325, 4352523, 4353201, 536 234, 935 47, 530351, C12N 1524, C12N 1563, C07K 1454

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061435244

ABSTRACT:
Provided is a fusion molecule comprising a DNA sequence encoding a thioredoxin-like protein fused to a DNA sequence encoding a second 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 the molecule. The fusion molecule may be modified to introduce one or more metal-binding/chelating amino-acid residues to aid in purification. 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.

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