KR940004051A - N-글리코실화에 결함이 있는 효모 숙주 균주 - Google Patents

N-글리코실화에 결함이 있는 효모 숙주 균주 Download PDF

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KR940004051A
KR940004051A KR1019930015252A KR930015252A KR940004051A KR 940004051 A KR940004051 A KR 940004051A KR 1019930015252 A KR1019930015252 A KR 1019930015252A KR 930015252 A KR930015252 A KR 930015252A KR 940004051 A KR940004051 A KR 940004051A
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레흐레 루드빅
레너르트 클라우스
코페츠키 에르하르트
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콜프, 융
뵈링거 만하임 게엠베하
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Abstract

본 발명은 〔3H〕-만노오스 자멸선택, 하나 또는 여러개의 선택적 마아커의 도입, 플라스미드 YEpL/GOD를 이용한 형질전환 및 표준조건하에 배양된 후 배양배자로 10㎖/ℓ 또는 그 이상의 GOD를 분리하고, 맥주효모 DSM7042, DSM 7338, DSM 7160 및/또는 DSM 7340의 ngd 돌연변이에 대립성이고 균일한 탄수화물 구조를 갖는 단백질을 분비하는 균주들의 선택에 의해 얻을 수 있는 N-글리코실화에 결함이 있는 효모 균주에 관한 것이다.

Description

Ｎ-글리코실화에 결함이 있는 효모 숙주 균주 [도면의 간단한 설명]
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 α-글루코시다아제 벡터인 XEp/5c6b3(12466 염시쌍)의 제한지도이고,
제2도는 맥주효모군에서 아스페르킬루스 니거 GOD의 분비를 위한 플라스미드를제조하기 위한 출발벡터인 XEpL(10717염기쌍;효모 발현벡터)의 제한지도이며,
제3도는 본 발명의 플라스미드 YEpL/GOD(12528 염기쌍)의 제한지도이다.

Claims (7)

  1. 〔3H〕-만노오스 자멸선택, 하나 또는 여러개의 선택적 마아커(영양요구성 및/또는 내성)의 도입, 플라스미드 YEpL/GOD를 이용한 형질전환 및 2% 효모추출물, 4% 박트펩톤(Difco), 0.1mod/ℓ 인산염완충액, pH7.0, 1% 프룩토오스 및 6%말토오스를 포함하는 완전배지에서 발효된 후 진동되면서 3일 내지 4일동안 인큐베이션된 후 배지로 10㎖/ℓ 또는 그 이상의 GOD를 분리하고, 맥주효모균 DSM7042, DSM 7338, DSM 7160 및/또는 DSM 7340에 대립성인 균주들의 선택에 의해 얻을 수 있는 N-글리코실화에 결함이 있는 효모 균주.
  2. 효모돌이연변이체, DSM7042, DSM 7338, DSM 7160 및 DSM 7340.
  3. 〔3H〕-만노오스 자멸선택, 하나 또는 여러개의 선택적 마아커(영양요구성 및/또는 내성)의 도입, 플라스미드 YEpL/GOD를 이용한 형질전환 및 2% 효모추출물, 4% 박트펩톤(Difco), 0.1mod/ℓ 인산염완충액, pH7.0, 1% 프룩토오스 및 6%말토오스를 포함하는 완전배지에서 발효된 후 진동되면서 3일 내지 4일동안 인큐베이션된 후 배지로 10㎖/ℓ 또는 그 이상의 GOD를 분리하고, 맥주효모균 DSM7042, DSM 7338, DSM 7160 및/또는 DSM 7340에 대립성인 균주들의 선택단계로 이루어지는 제1항 및 2항 기재의 효모 돌이변이체들의 제조방법.
  4. 제3항에 있어서, 맥주효모균 DSM7042, DSM 7338, DSM 7160 및/또는 DSM 7340의 제조를 위한 방법.
  5. 단백질의 동종 및 이종발현을 위한 숙주 유기체로서의 제1항 또는 2항의 표는 돌연변이체들의 용도.
  6. 원하는 단백질을 코드하는 재조합 DNA를 이용한 제1항 또는 제2항의 효모 돌연변이체의 형질전환, 세포의 발효 및 세로로부터 또는 배양상측액으로부터 원하는 단백질의 분리에 의해 얻을 수 있는 저글리코실화 재조합 단백질.
  7. 제1항 또는 2항의 효모 돌연변이체로부터의 단백질을 코드하는 DNA의 형질변환, 발효 및 세포 또는 배양 상층액으로부터 단백질의 분리에 의해 균일하게 글리코실화된 단백질의 발현 및 제조방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임
KR1019930015252A 1992-08-07 1993-08-06 N-글리코실화에 결함이 있는 효모 숙주 균주 KR0140973B1 (ko)

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DE4226094 1992-08-07
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DE4301932A DE4301932A1 (de) 1992-08-07 1993-01-25 Hefewirtsstämme mit Defekten in der N-Glycosylierung

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KR101460561B1 (ko) * 2012-11-20 2014-11-13 한국식품연구원 사카로마이세스 세레비지애 113-8의 대량 생산방법

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FR2750434B1 (fr) * 1996-06-26 1998-08-21 Applic De Rech Et De Conseils Produit de stabilisation proteique des vins
US6566122B1 (en) 2000-06-16 2003-05-20 Academia Sinica Super-secreting saccharomyces cerevisiae strains
WO2002000856A2 (en) * 2000-06-30 2002-01-03 Flanders Interuniversity Institute For Biotechnology (Vib) Protein glycosylation modification in pichia pastoris
KR100473658B1 (ko) * 2002-10-25 2005-03-10 드림바이오젠 주식회사 위치 선택적 수식법에 의한 혼성형 단백질 치료제 개발을 위한 형질전환 효모 시스템
US7507573B2 (en) 2003-11-14 2009-03-24 Vib, Vzw Modification of protein glycosylation in methylotrophic yeast
EP1934605B1 (en) * 2005-09-22 2014-03-26 Prosci Incorporated Glycosylated polypeptides produced in yeast mutants and methods of use thereof
WO2017170418A1 (ja) * 2016-03-30 2017-10-05 株式会社カネカ エンドヌクレアーゼ活性を有するポリペプチド及びその製造方法
CN107988177B (zh) * 2016-10-26 2021-08-20 潍坊康地恩生物科技有限公司 葡萄糖氧化酶突变体
CN108118037B (zh) * 2016-11-28 2021-08-31 青岛蔚蓝生物集团有限公司 一种耐热性提高的葡萄糖氧化酶突变体

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EP0314096B1 (en) * 1987-10-29 1994-01-12 Zymogenetics, Inc. Methods of regulating protein glycosylation
US5135854A (en) * 1987-10-29 1992-08-04 Zymogenetics, Inc. Methods of regulating protein glycosylation
NZ229260A (en) * 1988-06-03 1994-02-25 Merck & Co Inc Hepatitis b virus, expression cassette for pre-s domain, host cells and
US5094951A (en) * 1988-06-21 1992-03-10 Chiron Corporation Production of glucose oxidase in recombinant systems
EP0382332A3 (en) * 1989-01-06 1990-11-14 Collaborative Research Inc. Improved supersecreting mutants of saccharomyces cerevisiae
CA2022109A1 (en) * 1989-07-31 1991-02-01 Cheryl A. Schulman Enhanced expression of heterologous proteins in recombinant hosts in a non-growth media
DE4301904A1 (de) * 1992-08-07 1994-02-10 Boehringer Mannheim Gmbh Hypoglycosylierte recombinante Glucoseoxidase

Cited By (1)

* Cited by examiner, † Cited by third party
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KR101460561B1 (ko) * 2012-11-20 2014-11-13 한국식품연구원 사카로마이세스 세레비지애 113-8의 대량 생산방법

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DE4301932A1 (de) 1994-02-10
NO312246B1 (no) 2002-04-15
IL106589A (en) 2005-07-25
IL106589A0 (en) 1993-12-08
CA2103522A1 (en) 1994-02-08
NO932811L (no) 1994-02-08
US5705616A (en) 1998-01-06
NZ248323A (en) 1994-12-22
FI110006B (fi) 2002-11-15
ATE207955T1 (de) 2001-11-15
JPH06296482A (ja) 1994-10-25
EP0582244A3 (de) 1995-03-15
CA2103522C (en) 1998-07-07
EP0582244A2 (de) 1994-02-09
NO932811D0 (no) 1993-08-06
JP2984229B2 (ja) 1999-11-29
DE59310233D1 (de) 2001-12-06
EP0582244B1 (de) 2001-10-31
US5798226A (en) 1998-08-25
FI933487A (fi) 1994-02-08
AU657230B2 (en) 1995-03-02
FI933487A0 (fi) 1993-08-06
JPH1052277A (ja) 1998-02-24
KR0140973B1 (ko) 1998-06-15
AU4435493A (en) 1994-02-24
JP2675511B2 (ja) 1997-11-12

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