Horseshoe crab factor b variant
AssigneeSeikagaku Corporation
InventorsYuki KOBAYASHI, Hikaru MIZUMURA, Toshio ODA, Shun-ichiro KAWABATA
Life of this patent
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Abstract
Provided is a technology related to a horseshoe crab factor B variant, and also provided is means for performing endotoxin measurement with high sensitivity. A polypeptide having an amino acid sequence in which the amino acid residue at the 193-position in an amino acid sequence of a polypeptide of horseshoe crab factor B is substituted with a cysteine (Cys) residue, is produced. Endotoxin measurement can be carried out with high sensitivity by configuring a Limulus reagent by combining this polypeptide with horseshoe crab factor C.
Claims11
A polypeptide having an amino acid sequence in which the amino acid residue at the 193-position in an amino acid sequence of a polypeptide of horseshoe crab factor B is substituted with a cysteine (Cys) residue.
A polypeptide represented by any one of the following (A) to (D):
- (A) a polypeptide having an amino acid sequence represented by the following (A1) or (A2):
- (A1) an amino acid sequence represented by amino acid numbers 1 to 400 of SEQ ID NO:
- 7;
- and
- (A2) an amino acid sequence represented by amino acid numbers 24 to 400 of SEQ ID NO:
- 7,
- (B) a polypeptide having an amino acid sequence represented by the following (B1) or (B2):
- (B1) an amino acid sequence represented by amino acid numbers 1 to 400 of SEQ ID NO:
- 10;
- and
- (B2) an amino acid sequence represented by amino acid numbers 24 to 400 of SEQ ID NO:
- 10,
- (C) a polypeptide having an amino acid sequence including substitution, deletion, insertion, and/or addition of one amino acid residue or a plurality of amino acid residues in the amino acid sequence of the polypeptide represented by the item (A) or (B) (provided that the cysteine (Cys) residue at the 193-position is neither substituted nor deleted), the polypeptide having the function of horseshoe crab factor B, and
- (D) a polypeptide having an amino acid sequence of a fusion polypeptide in which a peptide tag is added to the polypeptide represented by any one of the items (A) to (C), the polypeptide having the function of horseshoe crab factor B.
A DNA represented by any one of the following (a) to (d):
- (a) a DNA having a base sequence represented by any one of the following (a1) to (a4):
- (a1) a base sequence represented by base numbers 1 to 1200 of SEQ ID NO:
- 5;
- (a2) a base sequence represented by base numbers 70 to 1200 of SEQ ID NO:
- 5;
- (a3) a base sequence represented by base numbers 1 to 1200 of SEQ ID NO:
- 6;
- and
- (a4) a base sequence represented by base numbers 70 to 1200 of SEQ ID NO:
- 6,
- (b) a DNA having a base sequence represented by any one of the following (b1) to (b4):
- (b1) a base sequence represented by base numbers 1 to 1200 of SEQ ID NO:
- 8;
- (b2) a base sequence represented by base numbers 70 to 1200 of SEQ ID NO:
- 8;
- (b3) a base sequence represented by base numbers 1 to 1200 of SEQ ID NO:
- 9;
- and
- (b4) a base sequence represented by base numbers 70 to 1200 of SEQ ID NO:
- 9,
- (c) a DNA hybridizing with a DNA comprising a base sequence complementary to the DNA represented by the item (a) or (b) under stringent conditions (provided that the bases represented by base numbers 577 to 579 are conserved), the DNA encoding a polypeptide having the function of horseshoe crab factor B, and
- (d) a DNA having the base sequence of a fusion DNA in which a peptide tag-encoding DNA is added to the DNA represented by any one of the items (a) to (c), the DNA encoding a polypeptide having the function of horseshoe crab factor B.
Dependencies are inferred from the claim text.