WO2001031023A1 - Nouveau polypeptide, proteine precurseur humaine 25 de la nucleoproteine ii, et polynucleotide codant pour ce polypeptide - Google Patents

Nouveau polypeptide, proteine precurseur humaine 25 de la nucleoproteine ii, et polynucleotide codant pour ce polypeptide Download PDF

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Publication number
WO2001031023A1
WO2001031023A1 PCT/CN2000/000380 CN0000380W WO0131023A1 WO 2001031023 A1 WO2001031023 A1 WO 2001031023A1 CN 0000380 W CN0000380 W CN 0000380W WO 0131023 A1 WO0131023 A1 WO 0131023A1
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WO
WIPO (PCT)
Prior art keywords
polypeptide
polynucleotide
precursor protein
human
nucleoprotein
Prior art date
Application number
PCT/CN2000/000380
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English (en)
Chinese (zh)
Inventor
Yumin Mao
Yi Xie
Original Assignee
Shanghai Bio Door Gene Technology Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Bio Door Gene Technology Ltd. filed Critical Shanghai Bio Door Gene Technology Ltd.
Priority to AU12645/01A priority Critical patent/AU1264501A/en
Publication of WO2001031023A1 publication Critical patent/WO2001031023A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy

Definitions

  • a polynucleotide encoding these isolated polypeptides, the polynucleotide comprising a nucleotide sequence having at least 91 nucleotides with a nucleotide sequence selected from the group consisting of »/» Identities: (a) a polynucleotide encoding the aforementioned human nucleoprotein II precursor protein 25; (b) a polynucleotide complementary to the polynucleotide (a).
  • the polynucleotide encodes a polypeptide having the amino acid sequence shown in SEQ ID NO: 2.
  • isolated human nucleoprotein ⁇ precursor protein 25 refers to human nucleoprotein I I precursor protein 25 is substantially free of other proteins, lipids, carbohydrates, or other substances naturally associated with it. Those skilled in the art can purify human nucleoprotein ⁇ precursor protein 25 using standard protein purification techniques. Substantially pure peptides can produce a single main band on a non-reducing polyacrylamide gel. The purity of the human nucleoprotein ⁇ precursor protein 25 can be analyzed by amino acid sequence.
  • a fragment, derivative or analog of the polypeptide of the present invention may be: U) a type in which one or more amino acid residues are replaced with conservative or non-conservative amino acid residues (preferably conservative amino acid residues), and the substituted
  • the amino acid may or may not be encoded by the genetic code; or ( ⁇ ) such that a group on one or more amino acid residues is substituted by another group to include a substituent; or ( ⁇ ⁇ )
  • One, in which the mature polypeptide is fused to another compound such as a compound that prolongs the half-life of the polypeptide, such as polyethylene glycol); or (IV) such a polypeptide sequence in which the additional amino acid sequence is fused into the mature polypeptide (such as the leader sequence or secreted sequence or the sequence used to purify this polypeptide or protease sequence)
  • the leader sequence or secreted sequence or the sequence used to purify this polypeptide or protease sequence such fragments, derivatives and analogs are considered to be within the knowledge of those
  • the invention also relates to variants of the polynucleotides described above, which encode polypeptides or fragments, analogs and derivatives of polypeptides having the same amino acid sequence as the invention.
  • This polynucleotide variant can be a naturally occurring allelic variant or a non-naturally occurring variant.
  • These nucleotide variants include substitution variants, deletion variants, and insertion variants.
  • an allelic variant is an alternative form of a polynucleotide that may be a substitution, deletion, or insertion of one or more nucleotides, but does not substantially change the function of the polypeptide it encodes .
  • “strict conditions” means: (1) hybridization and elution at lower ionic strength and higher temperature, such as 0.2xSSC, 0.1% SDS, 60 ° C; or (2) Add denaturants during hybridization, such as 50% (v / v) formamide, 0.1% calf serum / 0.1% Ficol 1, 42 ° C, etc .; or (3) only between two sequences Crosses occur only when the identity is at least 95%, and more preferably 97%.
  • the polypeptide encoded by the hybridizable polynucleotide has the same biological function and activity as the mature polypeptide shown in SEQ ID NO: 2.
  • Methods known to those skilled in the art can be used to construct expression vectors containing a DNA sequence encoding human nucleoprotein II precursor protein 25 and appropriate transcriptional / translational regulatory elements. These methods include in vitro recombinant DNA technology, DNA synthesis technology, in vivo recombination technology, etc. (Sambroook, et al. Mol ecular Cloning, a Laboratory Manua 1, cold Spring Harbor Laboratory. New York, 1989).
  • the DNA sequence can be operably linked to an appropriate promoter in an expression vector to guide mRNA synthesis. Representative examples of these promoters are: the lac or trp promoter of E.
  • polypeptide of the present invention can also be used for peptide mapping analysis.
  • the polypeptide can be specifically cleaved by physical, chemical or enzymatic analysis, and subjected to one-dimensional or two-dimensional or three-dimensional gel electrophoresis analysis, and more preferably mass spectrometry analysis.
  • the recombinant gene therapy vector can be used to treat diseases caused by abnormal expression or activity of human nuclear protein II precursor protein 25.
  • Virus-derived expression vectors such as retroviruses, adenoviruses, adenovirus-associated viruses, herpes simplex virus, parvoviruses, and the like can be used to transfer polynucleotides encoding human nucleoprotein ⁇ precursor protein 25 into cells.
  • a method for constructing a recombinant viral vector carrying a polynucleotide encoding a human nucleoprotein II precursor protein 25 can be found in the existing literature (Sarabrook, et al.).
  • a polynucleotide encoding human nucleoprotein ⁇ precursor protein 25 can be packaged into liposomes and transferred into cells.
  • a 32P-labeled probe (about 2 x 10 6 cpm / ml) was hybridized with a nitrocellulose membrane to which RNA was transferred at 42 ° C overnight in a solution containing 50% formamide-25mM KH 2 P0 4 (pH7.4) -5 530-5 06111 & ⁇ 1,3 solution and 20 ( ⁇ [111 salmon sperm 0. After hybridization, the filter was washed in 1 X SSC-0.1% SDS at 55 ° C for 30min. Then, Analysis and quantification using Phosphor Imager.
  • Example 5 In vitro expression, isolation and purification of recombinant human nucleoprotein II precursor protein 25

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Abstract

L'invention concerne un nouveau polypeptide, une protéine précurseur humaine 25 de la nucléoprotéine II, et un polynucléotide codant pour ce polypeptide ainsi qu'un procédé d'obtention de ce polypeptide par des techniques recombinantes d'ADN. L'invention concerne en outre les applications de ce polypeptide dans le traitement de maladies, notamment des tumeurs malignes, de l'hémopathie, de l'infection par VIH, de maladies immunitaires et de diverses inflammations. L'invention concerne aussi l'antagoniste agissant contre le polypeptide et son action thérapeutique ainsi que les applications de ce polynucléotide codant pour la protéine précurseur humaine 25 de la nucléoprotéine II.
PCT/CN2000/000380 1999-10-28 2000-10-27 Nouveau polypeptide, proteine precurseur humaine 25 de la nucleoproteine ii, et polynucleotide codant pour ce polypeptide WO2001031023A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU12645/01A AU1264501A (en) 1999-10-28 2000-10-27 A novel polypeptide-human nucleoprotein ii precursor protein 25 and the polynucleotide encoding said polypeptide

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN99119894.8 1999-10-28
CN 99119894 CN1302877A (zh) 1999-10-28 1999-10-28 一种新的多肽——人核蛋白ii前体蛋白25和编码这种多肽的多核苷酸

Publications (1)

Publication Number Publication Date
WO2001031023A1 true WO2001031023A1 (fr) 2001-05-03

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2000/000380 WO2001031023A1 (fr) 1999-10-28 2000-10-27 Nouveau polypeptide, proteine precurseur humaine 25 de la nucleoproteine ii, et polynucleotide codant pour ce polypeptide

Country Status (3)

Country Link
CN (1) CN1302877A (fr)
AU (1) AU1264501A (fr)
WO (1) WO2001031023A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100417726C (zh) * 2006-03-08 2008-09-10 哈尔滨工业大学 人源参与微丝组装调控的核蛋白及其制备和应用方法

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE EMBL [online] "BTCYTBC12", Database accession no. X59693 *
DATABASE GENBANK [online] 27 April 1993 (1993-04-27), "HUMCOR2M", Database accession no. J04973 *
DATABASE GENBANK [online] Database accession no. AC004069 *

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Publication number Publication date
CN1302877A (zh) 2001-07-11
AU1264501A (en) 2001-05-08

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