Process for production of recombinant proteins as a soluble form
AssigneeJnc Corporation
InventorsSatoshi Inouye, Yuiko Sahara
Life of this patent
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Abstract
A target protein is prepared as soluble protein using a recombinant protein expression system. An expression vector is used that includes (1) an expression-inducible promoter sequence; (2) a first coding sequence including a polynucleotide coding for a polypeptide that is represented by the formula (Z)n; and (3) a second coding sequence that includes a polynucleotide that codes for a target protein. A method of producing the target protein is also used that includes expressing protein using this expression vector.
Claims6
A fusion protein that can be expressed as a soluble protein, comprising:
- (1) a first amino acid sequence comprising the amino acid sequence of a polypeptide that is represented by the formula (Z)n,
- wherein n represents an integer from 1 to 5 and Z represents polypeptide selected from the group of:
- (a) a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1,
- (b) a polypeptide comprising an amino acid sequence that has at least 95% identity with the amino acid sequence of SEQ ID NO:
- 1, and
- (c) a polypeptide comprising an amino acid sequence that has at least 98% identity with the amino acid sequence of SEQ ID NO:
- 1,
- and that has the capacity when expressed as a fusion protein with a target protein to render said fusion protein expressible as a soluble protein;
- and
- (2) a second amino acid sequence comprising the amino acid sequence of a target protein,
- wherein the fusion protein further comprises, on the amino terminal side of the first amino acid sequence, an amino acid sequence that facilitates purification, and;
- wherein the expression of the fusion protein comprising the amino acid sequence that facilitates purification, an amino acid sequence of the polypeptide that is represented by the formula (Z)n and an amino acid sequence of the target protein is induced in Escherichia coli, and the fusion protein accumulates within the Escherichia coli as a soluble protein so that the fusion protein collected from within the Escherichia coli has higher solubility rate than non-(Z)n fusion protein;
- and
- wherein the amino acid sequence that facilitates the purification is a histidine tag sequence;
- and
- wherein the target protein is at least one selected from the group of apoaequorin, Gaussia luciferase, and Oplophorus luciferase.
A fusion protein that can be expressed as a soluble protein and that is represented by the formula (Z)n-L-X,
- wherein n represents an integer from 1 to 5;
- L represents a cleavable linker peptide;
- Z represents polypeptide selected from the group of:
- (a) a polypeptide comprising the amino acid sequence of SEQ ID NO:
- 1,
- (b) a polypeptide comprising an amino acid sequence that has at least 95% identity with the amino acid sequence of SEQ ID NO:
- 1, and
- (c) a polypeptide comprising an amino acid sequence that has at least 98% identity with the amino acid sequence of SEQ ID NO:
- 1, and
- X represents the amino acid sequence of a target protein,
- wherein the fusion protein further comprises, on the amino terminal side of Z, an amino acid sequence that facilitates purification, and;
- wherein the expression of the fusion protein is induced in Escherichia coli, and the fusion protein accumulates within the Escherichia coli as a soluble protein so that the fusion protein collected from within the Escherichia coli has higher solubility rate than non-(Z)n fusion protein;
- and
- wherein the amino acid sequence that facilitates the purification is a histidine tag sequence;
- and
- wherein the target protein is at least one selected from the group of apoaequorin, Gaussia luciferase, and Oplophorus luciferase.
Dependencies are inferred from the claim text.