IP Library Granted Patent US 12,371,697
Granted Patent B2
US 12,371,697 · App. 17/018,751 · Granted Jul 29, 2025

Peptide tag and tagged protein including same

Inventors: Kazuyoshi Koike (Sodegaura, JP); Eiji Takita (Sodegaura, JP); Koichi Kaneda (Sodegaura, JP)
Assignee: IDEMITSU KOSAN CO., LTD
C12N15/62C07K4/00C07K7/06C07K7/08C07K14/00C07K14/39C07K14/43504C07K14/43563C07K14/47C07K14/475C07K14/565C07K19/00C12N5/10C12N9/248C12N15/09C12P21/00C12P21/02C12Q1/6806C07K2319/40C07K2319/50C07K2319/60C12Q2531/113C12Y302/01
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,371,697
App. No.
17/018,751
Granted
Jul 29, 2025
Kind
B2
Abstract

A peptide comprising the sequence shown below is added as a peptide tag to a useful protein, followed by allowing its expression. X m (PY n ) q PZ r In this formula, X, Y, and Z each represent an amino acid residue independently selected from the group consisting of R, G, S, K, T, L, N, Q, and H, with the proviso that at least one Y represents K, L, N, Q, H, or R. m represents an integer of 0 to 5; n represents 1, 2, or 3; q represents an integer of 1 to 10; and r represents an integer of 0 to 10.

Claims (21)

1. A DNA encoding a tagged protein, comprising:

a peptide; and

a useful protein,

wherein the peptide consists of sequence X m (PY n ) q PZ r , where X and Z each represent an amino acid residue independently selected from the group consisting of arginine (R), glycine (G), serine (S), threonine (T), leucine (L), asparagine (N), glutamine (Q), and histidine (H), and Y represents an amino acid residue independently selected from the group consisting of glycine (G), serine (S), lysine (K), threonine (T), leucine (L), asparagine (N), glutamine (Q), and histidine (H), with the proviso that at least one X is R and at least one Y in each PY n represents K; m represents an integer of 1 to 5; n represents 2, or 3; q represents an integer of 2 to 10; and r represents an integer of 1 to 10, and the peptide consists of 9 to 50 amino acids.

2. A recombinant vector comprising the DNA according to claim 1 .

3. A transformant prepared by transformation with the DNA according to claim 1 .

4. A transformant prepared by transformation with the recombinant vector according to claim 2 .

5. The transformant according to claim 3 , wherein said transformant is yeast, E. coli, Brevibacillus , an insect cell, or a mammalian cell, and wherein the mammalian cell comprises a human cultured cell, but does not comprise a human individual.

6. The transformant according to claim 4 , wherein said transformant is yeast, E. coli, Brevibacillus , an insect cell, or a mammalian cell, and wherein the mammalian cell comprises a human cultured cell, but does not comprise a human individual.

7. A method for producing a tagged protein, the method comprising:

culturing the transformant according to claim 3 to allow accumulation of the tagged protein, and

collecting the tagged protein.

8. A method for producing a tagged protein, the method comprising:

culturing the transformant according to claim 4 to allow accumulation of the tagged protein, and

collecting the tagged protein.

9. The DNA according to claim 1 , wherein the content of G and S of said peptide is less than 60%.

10. A DNA encoding a tagged protein, wherein said tagged protein comprises a peptide and a useful protein, the peptide comprises the amino acid sequence of SEQ ID NO: 25, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 58, 60, 64, 66, 92, 94, 96, 98, 100, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 140, 142, 143, 145, 147, 149, 151, 153, 155, or 157.

11. The DNA according to claim 1 , wherein said peptide is bound to the N-terminus, the C-terminus or to each of both the N-terminus and the C-terminus of the useful protein.

12. The DNA according to claim 1 , wherein said useful protein is selected from the group consisting of human growth hormone, interferon β, xylanase, esterase, and green fluorescent protein (GFP).

13. The DNA according to claim 1 , wherein said peptide is linked to the useful protein through a protease recognition sequence.

14. The DNA according to claim 1 , wherein said tagged protein further comprising a secretion signal.

Priority Claims (2)
JP 2015-256396 · Dec 28, 2015 · national
JP 2016-153265 · Aug 3, 2016 · national
Continuity (2)
Continuation 16066468
Related Publication 20210130404A1 · May 6, 2021
References Cited (45)
US 6686333B1 · Kashanchi · 2004 [cited by applicant]
US 7205387B2 · Wang · 2007 [cited by examiner]
US 20020098524A1 · Murray · 2002 [cited by examiner]
US 20090137004A1 · Uversky et al. · 2009 [cited by applicant]
US 20100234568A1 · Decarolis · 2010 [cited by examiner]
US 20110033389A1 · Chen · 2011 [cited by examiner]
US 20110041220A1 · Ohto et al. · 2011 [cited by applicant]
US 20110231960A1 · Sawada et al. · 2011 [cited by applicant]
US 20140329742A1 · Dock · 2014 [cited by applicant]
CA 2765171 · 2010 [cited by applicant]
JP 2009201437A · 2009 [cited by applicant]
JP 2010226992A · 2010 [cited by examiner]
JP 5273438B2 · 2013 [cited by applicant]
JP 5360727B2 · 2013 [cited by applicant]
JP 2014520507A · 2014 [cited by applicant]
JP 5626717B2 · 2014 [cited by examiner]
WO WO2009133882A1 · 2009 [cited by applicant]
WO WO2010129033A2 · 2010 [cited by examiner]
WO WO2013091661A2 · 2013 [cited by examiner]
WO WO2013130683A2 · 2013 [cited by examiner]
WO WO2014036562A2 · 2014 [cited by applicant]
Choi et al. “Secretory and extracellular production of recombinant protein using [cited by examiner]
Low et al. “Optimisation of signal peptide for recombinant protein secretion in bacterial hosts”, Appl Microbiol Biotechnol, 2013, 3811-3826 (Year: 2013). [cited by examiner]
Expasy-PeptideCutter, https://web.expasy.org/peptide_cutter/, accessed on Oct. 20, 2023 (Year: 2023). [cited by examiner]
Snapp “Design and Used of Fluorescent Fusion Proteins in Cell Biology”, Curr Protoc Cell Biol, 2005, Unit 21.4 (Year: 2005). [cited by examiner]
Weiss et al., “Intracellular peptide delivery using amphiphilic lipid-based formulations”, Biotechnology and Bioengineering, 2477-2487 (Year: 2011). [cited by examiner]
Decision of Refusal issued Sep. 28, 2021 in Japanese Patent Application No. 2017-559238 (with English machine translation), 9 pages. [cited by applicant]
Japanese Office Action issued Jan. 5, 2021 in Japanese Patent Application No. 2017-559238 (with English translation), 10 pages. [cited by applicant]
International Search Report issued Apr. 4, 2017 in PCT/JP2016/089139, 2 pages. [cited by applicant]
International Preliminary Report on Patentability and Written Opinion issued Jul. 12, 2018 in PCT/JP2016/089139 (submitting English translation only), 12 pages. [cited by applicant]
Nierman, W.C., et al., Accession No. Q9AB24, Definition: TPR Domain Protein, Jun. 1, 2001, Retrieved from the internet: URL: http://www.uniprot.org/uniprot/Q4AB24.txt?version=1 , 1 page. [cited by applicant]
Smith, D.B., et al., “Single-step purification of polypeptides expressed in [cited by applicant]
Marblestone, J.G., et al., “Comparison of SUMO fusion technology with traditional gene fusion systems: Enhanced expression and solubility with SUMO”, Protein Science. vol. 15, 2006, pp. 182-189. [cited by applicant]
Guan, C.D., et al., “Vectors that facilitate the expression and purification of foreign peptides in [cited by applicant]
Maina, C.V., et al., “An [cited by applicant]
Extended European Search Report issued May 22, 2019 in Patent Application No. 16881827.6, 8 pages. [cited by applicant]
Wu, D. et al. “High-level secretory expression of metchnikowin in [cited by applicant]
“Uncharacterized protein from Candida parapsilosis” Database UniProt, https://www.uniprot.org/uniorot/G8B5T8.txt, XP002791092, 2012, 2 Pages. [cited by applicant]
GenBank: CAN98496.1; conjugal transfer protein; 2016; pp. 1-2 (Year: 2016). [cited by applicant]
Uni ProtKB/Swiss-Prot: P17656.1; Cuticle collagen 2, 2006, pp. 1-2 (Year: 2006). [cited by applicant]
UniProtKB/Swiss-Prot: Q9HX66; Probable ATP-dependent RNA helicase, 2006, p. 1 (Year: 2006). [cited by applicant]
UniProtKB/Swiss-Prot: Q9AB24; TPR domain protein, 2006, p. 1 (Year: 2006). [cited by applicant]
Gen Bank: KDB11109.1; RNA polymerase i specific transcription initiation factor of the organism [ [cited by applicant]
Argiolas et al., “Amino Acid Sequence of Bumblebee MCD Peptide: A New Mast Cell Degranulating Peptide From the Venom of the Bumblebee [cited by applicant]
Office Action issued in corresponding Canadian Patent Application No. 3,009,880 on Nov. 22, 2023. [cited by applicant]