KR970703427A - 식물, 진균류 및 미생물에서 선형 α-1,4 글루칸의 합성을 용이하게 할 수 있는 효소를 코딩하는 DNA 서열(DNA SEQUENCES CODING FOR ENZYMES CAPABLE OF FACILITATING THE SYNTHESIS OF LINEAR α-1,4 GLUCANS IN PLANTS, FUNGI AND MICROORGANISMS) - Google Patents
식물, 진균류 및 미생물에서 선형 α-1,4 글루칸의 합성을 용이하게 할 수 있는 효소를 코딩하는 DNA 서열(DNA SEQUENCES CODING FOR ENZYMES CAPABLE OF FACILITATING THE SYNTHESIS OF LINEAR α-1,4 GLUCANS IN PLANTS, FUNGI AND MICROORGANISMS) Download PDFInfo
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Abstract
본 발명은 박테리아, 진균류 및 식물에 의하여 또는 세포-유리 시스템내에서, 자당으로부터 선형 α-1,4 글루칸을 합성시키는 아밀로수크라아제의 효소 활성을 갖는 단백질을 코딩하는 DNA 서열; 상기 DNA 서열을 함유하는 플라스미드 및 박테리아; 세포내 또는 세포외적으로 아밀로수크라아제 활성을 갖는 폴리펩티드를 발현시킬 수 있는 식물 및 미생물의 제조방법; 및 아밀로수크라아제의 효소 활성을 나타내는 단백질을 사용하여 순수한 과당을 제조하는 것에 관한 것으로서, 이중, 상기 DNA 서열은 하기와 같은 단계로 제조되는 것을 특징으로 한다.
(a) 생물체 세포의 게놈성 DNA 또는 mRNA를 기본으로 게놈성 또는 cDNA 라이브러리를 제조하고; (b) (a)에 따라 제조된 라이브러리로 적당한 숙주를 형질전환시키고; (c) 형질전환된 세포에 요오드 증기를 가하고; (d) 푸른색으로 착색된 세포를 규명하고; (e) (d)에서 규명된 세포를 분리 및 배양하고; (f) 형질전환된 세포로부터 게놈성 DNA 삽입체 또는 cDNA 삽입체를 분리함.
Description
식물, 진균류 및 미생물에서 선형 α-1,4 글루칸의 합성을 용이하게 할 수 있는 효소를 코딩하는 DNA 서열(DNA SEQUENCES CODING FOR ENZYMES CAPABLE OF FACILITATING THE SYNTHESIS OF LINEAR α-1,4 GLUCANS IN PLANTS, FUNGI AND MICROORGANISMS)
[도면의 간단한 설명]
제1도는 플라스미드 pNB2 (DSM 9196)을 보여주고 있다.
제2도는 요오드 증가를 가함으로써 형질전환된 E. coli 세포내 아밀로수크라아제 활성의 발현을 나타낸 것이다.
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
Claims (21)
- 하기로 구성된 군으로부터 선택되는 것을 특징으로 하는, 아밀로수크라아제의 효소 활성을 갖는 단백질을 코딩하는 DNA 서열.(a) 서열번호 1로 표시되는 아미노산 서열을 갖는 단백질을 코딩하는 DNA 서열; (b) 서열번호 1로 표시되는 코딩영역을 나타내는 DNA 서열; (c) (a) 또는 (b)의 서열 중 어느 하나에 하이브리드되는 DNA 서열; 및 (d) (a), (b) 또는 (c)에서 언급된 서열에 비교시 유전 코드에 의하여 변질된 DNA 서열.
- 하기 단계로 구성되는 방법에 의하여 수득될 수 있는 것을 특징으로 하는, 아밀로수크라아제의 효소 활성을 갖는 단백질을 코딩하는 DNA 서열 : (a) 생물체 세포의 게놈성 DNA 또는 mRNA를 기본으로 게놈성 또는 cDNA 라이브러리를 제조하고; (b) (a)에 따라 제조된 라이브러리로 적당한 숙주를 형질전환시키고; (c) 형질 전환된 세포에 요오드 증기를 가하고; (d) 푸른색으로 착색된 세포를 규명하고; (e) (d)에서 규명된 세포를 분리 및 배양하고; (f) 형질전환된 세포로부터 게놈성 DNA 삽입체 또는 cDNA 삽입체를 분리함.
- 제1항 또는 제2항에 따른 DNA 서열을 함유하는 것을 특징으로 하는 재조합 DNA 분자.
- 제3항에 있어서, 아밀로수크라아제의 효소 활성을 갖는 단백질을 코딩하는 DNA 서열을, 진핵 또는 원핵 세포내에서 전자시키는 DNA 서열과 연결시키는 것을 특징으로 하는 재조합 DNA 분자.
- DSM 9196으로 기탁된 플라스미드 pNB2.
- 제3항 내지 제5항 중 어느 한 항에 따른 재조합 DNA 분자를 함유하는 것을 특징으로 하는 미생물.
- 제3항 내지 제5항 중 어느 한 항에 따른 재조합 DNA 분자를 함유하는 것을 특징으로 하는 진균류.
- 제1항 또는 제2항에 다른 DNA 서열을 코딩하는 것을 특징으로 하는 아밀로수크라아제의 효소 활성을 갖는 단백질.
- 제6항에 따른 미생물 또는 제7항에 따른 진균류를 적당한 배양 배지내에서 배양하는 것을 특징으로 하는, 제8항에 따른 단백질의 제조 방법.
- 제1항 또는 제2항에 따른 DNA 서열을 발현시키는 DNA 서열에 연결된 DNA 서열이 식물 세포내로 도입된 후 전체 식물이 상기 식물 세포로부터 재생되는 것을 특징으로 하는, 선형 α-1,4 글루칸을 합성할 수 있는 식물의 제조 방법.
- 제10항에 있어서, 하기 단계로 이루어진 것을 특징으로 하는 방법; (a) 하기 부분적인 서열을 갖는 발현 카셋트를 제조하고; (i) 각각의 표적 조직 또는 표적 세포에서 RNA를 형성하게 하고 식물에서 활성이 있는 프로모터; (ii) 아밀로수크라아제의 효소 활성을 갖는 단백질을 코딩하며 센스 방향에서 프로모터에 융합되는, 제10항에서 설명된 것과 같은 적어도 하나의 DNA 서열; (iii) RNA 분자의 전사 종결 및 폴리아데닐화를 위한, 식물에서 기능성이 있는 신호; (b) 상기 발현 카셋트를 식물 세포내로 전달하고; (c) 형질전환된 식물 세포로부터 본래의 전체 식물을 재생함.
- 제1항 또는 제2항에 따른 DNA 서열이 미생물로 도입된 후 발현되는 것을 특징으로 하는, 선형 α-1,4 글루칸을 합성할 수 있는 미생물의 제조 방법.
- 제12항에 있어서, 하기 단계로 이루어진 것을 특징으로 하는 방법; (a) 하기 부분적인 서열을 갖는 발현 카셋트를 제조하고; (i) 선택된 미생물에서 활성이 있으며, DNA 서열 다운스트림을 전사시키는 프로모터; (ii) 아밀로수크라아제를 코딩하며, 센스방향으로 상기 프로모터에 융합되어 있는 DNA 서열; (iii) 미생물에서 기능이 있는 전사 종결 신호; 및 (b) 단계 (a)에서 제조된 발현 카셋트로 적당한 미생물을 형질전환시킴.
- 제1항 또는 제2항에 따른 DNA 서열이 진균 세포내로 도입된 후 발현되는 것을 특징으로 하는, 선형 α-1,4 글루칸을 합성할 수 있는 진균 세포의 제조 방법.
- 제14항에 있어서, 하기 단계로 구성된 것을 특징으로 하는 방법 : (a) 하기 부분적인 서열을 갖는 발현 카셋트를 제조하고 : (i) 선택된 진균류의 세포에서 활성이 있으며, DNA 서열 다운스트림을 전사시키는 프로모터; (ii) 아밀로수크라아제를 코딩하며, 센스방향으로 상기 프로모터에 융합되어 있는 DNA 서열; (iii) 진균류 세포에서 기능이 있는 전사 종결 신호; 및 (b) 단계 (a)에서 제조된 발현 카셋트로 진균류 세포를 형질전환시킴.
- 식물세포에서 DNA 서열을 발현시키는 DNA 서열과 결합되어, 제1항 또는 제2항에 따른 DNA 서열을 함유하는 것을 특징으로 하는 유전전환성 식물 세포 및 식물.
- 제16항에 있어서, 상기 식물이 곡류인 것을 특징으로 하는 식물.
- 제16항 또는 제17항에 있어서, 상기 식물이 옥수수, 벼, 밀, 보리, 사탕무우, 사탕수수, 담배, 토마토, 또는 감자인 것을 특징으로 하는 식물.
- 제1항 또는 제2항에 따른 DNA 서열 또는 상동의 DNA 또는 RNA 서열을 분리하기 위하여, 그로부터 유래된 프로브 분자를 사용하는 것을 특징으로 하는 상기 DNA 서열 또는 프로브 분자의 용도.
- 선형 α-1,4 글루칸의 제조시 사용되는 것을 특징으로 하는 제8항에 따른 단백질의 용도.
- 과당의 제조시 사용되는 것을 특징으로 하는 제8항에 따른 단백질의 용도.※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
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DE19944417879 DE4417879A1 (de) | 1994-05-18 | 1994-05-18 | DNA-Sequenzen die die Synthese linearer alpha-1,4-Glukane in Pflanzen, Pilzen und Mikroorganismen ermöglichen |
DEP4417879.4 | 1994-05-18 | ||
DEP4447388.5 | 1994-12-22 | ||
DE19944447388 DE4447388A1 (de) | 1994-12-22 | 1994-12-22 | DNA-Sequenzen, die die Synthese linearer alpha-1,4-Glukane in Pflanzen, Pilzen und Mikroorganismen ermöglichen |
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KR970703427A true KR970703427A (ko) | 1997-07-03 |
KR100352532B1 KR100352532B1 (ko) | 2004-02-18 |
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AU3852089A (en) | 1988-06-21 | 1990-01-12 | Calgene, Inc. | Methods and compositions for altering physical characteristics of fruit and fruit products |
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1995
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- 1995-05-18 PT PT95920833T patent/PT759993E/pt unknown
- 1995-05-18 US US08/737,752 patent/US6265635B1/en not_active Expired - Lifetime
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- 1995-05-18 KR KR1019960706453A patent/KR100352532B1/ko not_active IP Right Cessation
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- 1995-05-18 DK DK95920833T patent/DK0759993T3/da active
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AU2614195A (en) | 1995-12-05 |
DK0759993T3 (da) | 2007-11-12 |
HUT76087A (en) | 1997-06-30 |
HU9603170D0 (en) | 1997-01-28 |
AU699552B2 (en) | 1998-12-10 |
IL113776A0 (en) | 1995-08-31 |
ATE368118T1 (de) | 2007-08-15 |
RU2201963C2 (ru) | 2003-04-10 |
DE69535543T2 (de) | 2008-04-30 |
DE69535543D1 (de) | 2007-09-06 |
JPH10500297A (ja) | 1998-01-13 |
US20050227319A1 (en) | 2005-10-13 |
CA2190149A1 (en) | 1995-11-23 |
HU226874B1 (en) | 2010-01-28 |
US7456003B2 (en) | 2008-11-25 |
JP2003325067A (ja) | 2003-11-18 |
CA2190149C (en) | 2011-01-04 |
IL113776A (en) | 2008-12-29 |
PT759993E (pt) | 2007-08-10 |
ES2287935T3 (es) | 2007-12-16 |
US6265635B1 (en) | 2001-07-24 |
EP0759993B1 (en) | 2007-07-25 |
JP3555086B2 (ja) | 2004-08-18 |
JP3511027B2 (ja) | 2004-03-29 |
US6818421B2 (en) | 2004-11-16 |
WO1995031553A1 (en) | 1995-11-23 |
KR100352532B1 (ko) | 2004-02-18 |
EP0759993A1 (en) | 1997-03-05 |
US20020092040A1 (en) | 2002-07-11 |
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