KR900013077A - 글루카곤의 제조방법 - Google Patents

글루카곤의 제조방법 Download PDF

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KR900013077A
KR900013077A KR1019900001157A KR900001157A KR900013077A KR 900013077 A KR900013077 A KR 900013077A KR 1019900001157 A KR1019900001157 A KR 1019900001157A KR 900001157 A KR900001157 A KR 900001157A KR 900013077 A KR900013077 A KR 900013077A
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glucagon
fusion protein
trp
amino acid
glu
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KR1019900001157A
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KR0161656B1 (ko
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쥰 이시자키
미키오 다마키
마사루 신
히로시 데라오카
노브오 요시다
히로시게 쯔즈기
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원본 미기재
시오노기 세이야구 가부시기가이샤
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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
    • C07K14/52—Cytokines; Lymphokines; Interferons
    • C07K14/555—Interferons [IFN]
    • C07K14/57—IFN-gamma
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • 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
    • C07K14/575—Hormones
    • C07K14/605—Glucagons
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/50—Fusion polypeptide containing protease site
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/74—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor
    • C07K2319/75—Fusion polypeptide containing domain for protein-protein interaction containing a fusion for binding to a cell surface receptor containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones

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Abstract

내용 없음

Description

글루카곤의 제조방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 인간 글루카곤을 암호화하는 DNA서열을 나타낸다.
제2(a)도는 본 발명의 발현 벡터 Trp pAT huFNr/글루카곤의 제조에 사요오디는 플라스미드 pGA1의 제조를 나타내는 개략도이다.
제3도는 DNA의 상류 또는 하류에 인접한 부위 뿐만 아니라 발현벡터 Trp pAT huFNr/글루카곤을 함유하는 글루카곤을 암호화하는 DNA염기 서열을 나타낸다.

Claims (11)

1. 다음식(Ⅰ)의 표시되는 융합단백질을 제조하고
x―Glu―Glucagon (Ⅰ)
(여기에서, X는 70내지170개의 아미노산 리더 서열이고, Glu는 글루탐산 잔기이고, Glucagon은 글루카곤의 아미노산 서열을 나타낸다), 상기 융합 단백질을 글루탐산 잔기의 카복실 말단에서 펩티드결합을 특이적으로 가수분해하는 효소로 처리하여 융합 단백질을 절단하는 것을 포함하는 글루카곤의 제조방법.
제1항에 있어서, 서열 X가 인터페론―r, △ Cys 인터페론―r 또는 인터페론 α80A2의 아미노 말단 서열일부를 함유하는 방법.
제1항에 있어서, 글루카곤의 아미노산 서열이 다음식(Ⅱ)으로 표시되는 방법.
His Ser Gln Gly Thr Phe Thr Ser Asp Yyr
Ser Lys Tyr Leu Asp Ser Arg Arg Ala Gln
Asp Phe Val Gln Trp Leu Met Asn Thr (Ⅱ)
제1항에 있어서, 융합 단백질을 식(Ⅰ)로 표시되는 융합 단백질을 발현하는 발현 벡터를E. coli로 도입함으로써 얻어지는 형질전환체를 배양시키는 방법으로 제조하는 방법.
제1항에 있어서, 글루카곤을 (a)융합단백질을 적절한 변성화제 또는 계면활성제로 용해시키고, (b)변성제 또는 계면활성제의 농도를 저하시키고, (c)(b)단계에서 얻은 혼합물에 얻은 혼합물을에 효소를 가하여 융합 단백질의 부분 분해에서 생성되는 가용성 중간물질 함유한 반응 혼합물을 얻고 (d)반응혼합물을 탈염시켜서 중간물질 및 효소를 함유하고 변성제 또는 계면활성제는 함유하지 않는 획분을 얻고, (e)획분에 함유된 효소에 의해 반응을 더 진행시켜서 글루코겐을 방출시키고, (f)방출된 글루코겐을 분리시키는 것을 포함하는 단계에 의해 회수하는 방법.
제1항에 있어서, 효소가스타필로코쿠스 아우레우스로부터 유도된 프로테아제 V8인 방법.
다음식 (Ⅲ)으로 표시되는, 글루카곤을 암호화하는 DNA서열.
5'―CACTCTCAGGGTACCTTCACTTCCGACTACTCTAAA
TACCTGGACTCCCGTCGTGCTCAGGACTTCGTTCAG
TGGCTGATGAACACC―3' (Ⅲ)
다음식(Ⅰ)으로 표시되는 융합 단백질을 암호화하는 DNA서열을 포함하는 발현벡터
X―Glu―Glucagon (Ⅰ)
X는 70내지 170개의 아미노산 리더 서열이고, Glu는 글루탐산 잔기이고, Glucagon은 글루카곤의 아미노산 서열을 나타낸다.
제8항에 있어서, 발현벡터가 Trp pAT huIFN r/글루카곤, Trp pAT△Cys huIFN r/글루카곤, Trp pAT△Cys쇼트 huIFN r/글루카곤, Trp pAT huIFN r80A2/글루카곤 및 Trp pAT△CysⅡhuIFN r/글루카곤으로 구성된 그룹에서 선택된 발현 벡터.
제8항 또는 제9항의 발현 벡터를E. coli의 세포로 도입하여 얻는 형절전환체.
고성능 액체 크로마토그래프로 측정하여 99% 이상의 순도를 갖는 정제된 글루카곤.
※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019900001157A 1989-02-01 1990-02-01 글루카곤의 생산방법 KR0161656B1 (ko)

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JP89-24158 1989-02-01
JP1-24158 1989-02-01
JP2415889 1989-02-01

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KR900013077A true KR900013077A (ko) 1990-09-03
KR0161656B1 KR0161656B1 (ko) 1998-11-16

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EP (1) EP0381433B1 (ko)
JP (1) JPH0748999B2 (ko)
KR (1) KR0161656B1 (ko)
CN (1) CN1063795C (ko)
AT (1) ATE144551T1 (ko)
BR (1) BR9000445A (ko)
CA (1) CA2008824C (ko)
DE (1) DE69028935T2 (ko)
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IL (1) IL93173A0 (ko)
NO (1) NO900410L (ko)
NZ (1) NZ232255A (ko)
PT (1) PT93031A (ko)
ZA (1) ZA90533B (ko)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6506595B2 (en) * 1998-03-31 2003-01-14 Itoham Foods Inc. DNAs encoding new fusion proteins and processes for preparing useful polypeptides through expression of the DNAs
CN100480374C (zh) * 2001-09-28 2009-04-22 中国科学院上海生命科学研究院 制备重组人类胰高血糖素肽－1氨基酸7－37肽段的方法
BR112012024379A2 (pt) * 2010-03-26 2017-01-10 Novo Nordisk As "peptídeos glucagon, seu uso, bem como composição farmacêutica"
MX354705B (es) 2011-09-23 2018-03-16 Novo Nordisk As Analogos de glucagon novedosos.
MY174727A (en) 2013-04-18 2020-05-11 Novo Nordisk As Stable, protracted glp-1/glucagon receptor co-agonists for medical use
US10570184B2 (en) 2014-06-04 2020-02-25 Novo Nordisk A/S GLP-1/glucagon receptor co-agonists for medical use
CN104593399A (zh) * 2014-12-22 2015-05-06 广西大学 一种转基因胰岛素的合成工艺

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DK27885A (da) * 1985-01-22 1986-07-23 Novo Industri As Fremstilling af peptider
US4743679A (en) * 1986-02-24 1988-05-10 Creative Biomolecules, Inc. Process for producing human epidermal growth factor and analogs thereof

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DK0381433T3 (da) 1996-11-18
KR0161656B1 (ko) 1998-11-16
FI900497A0 (fi) 1990-01-31
PT93031A (pt) 1990-08-31
HU900614D0 (en) 1990-04-28
NO900410D0 (no) 1990-01-29
ZA90533B (en) 1990-10-31
ES2095236T3 (es) 1997-02-16
DE69028935D1 (de) 1996-11-28
CA2008824C (en) 1997-08-05
JPH0748999B2 (ja) 1995-05-31
CN1046559A (zh) 1990-10-31
EP0381433A1 (en) 1990-08-08
DE69028935T2 (de) 1997-03-06
NO900410L (no) 1990-08-02
GR3021965T3 (en) 1997-03-31
JPH03254692A (ja) 1991-11-13
EP0381433B1 (en) 1996-10-23
AU4898990A (en) 1990-08-09
CN1063795C (zh) 2001-03-28
NZ232255A (en) 1992-03-26
IL93173A0 (en) 1990-11-05
CA2008824A1 (en) 1990-08-01
ATE144551T1 (de) 1996-11-15
HUT54410A (en) 1991-02-28
BR9000445A (pt) 1991-01-15
AU634999B2 (en) 1993-03-11

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