KR930023461A - 안정성있는 크로모좀 유전자 증폭을 위한 간단한 방법 - Google Patents

안정성있는 크로모좀 유전자 증폭을 위한 간단한 방법 Download PDF

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KR930023461A
KR930023461A KR1019930007771A KR930007771A KR930023461A KR 930023461 A KR930023461 A KR 930023461A KR 1019930007771 A KR1019930007771 A KR 1019930007771A KR 930007771 A KR930007771 A KR 930007771A KR 930023461 A KR930023461 A KR 930023461A
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dna
dna sequence
polypeptide
encoding
host cell
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KR1019930007771A
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베터 로만
빌케 데트레프
쾰러 마그레트
아모리 안토이너
묄러 베른하르트
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닥터 라우어
솔베이 엔침스 게엠베하 운트 콤파니, 콤만디트게젤샤프트
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Publication of KR930023461A publication Critical patent/KR930023461A/ko

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    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
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    • C12Y302/01001—Alpha-amylase (3.2.1.1)

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Abstract

본 발명은, 특히 바실러스 종의 원핵 미생물에서 폴리펩타이드를 코트하는 DAN서열을 안정하게 크로모좀 유전자 증폭시키기 위한 간단한 방법에 관한 것이다.

Description

안정성있는 크로모좀 유전자 증폭을 위한 간단한 방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 플라스미드 pCLEAN4의 제한지도, 제2도는 플라스미드 pUB 131의 제한지도, 제3도는 플라스미드 pLI의 제한지도.

Claims (6)

  1. 해당 폴리펩타이드를 코드하는 DNA 서열의 적어도 한 카피를 그의 크로모좀 DNA 상에 이미 가지고 있는 원핵 숙주세포를 다음의 가)와 나)를 필수적으로 포함하며, 숙주세포내에서 자가복제할 수 있는 DNA서열을 포함하는 환형 DNA없이 숙주세포의 형질전환에 적합한 환형 DNA를 이용하여 형질전환시키며, 사용된 환영 DNA 적어도 한 카피에서 크로모좀 통합된 세포를 선택성이 있는 마-커의 도움으로 선별해 내고, 그렇게 형질전환된 세포를 분리하는 것을 특징으로 하는 해당 폴리펩타이드를 코드하는 DNA 서열의 적어도 2 카피를 크로모좀 상에 포함하고 있는 형질전환된 원핵 숙주세포의 제조방법.
    가)해당 폴리펩타이드의 발현과 필요한 경우 분비에 필요한 서열을 포함하는 해당 폴리펩타이드를 코드하는 DNA 서열을 포함하는 DNA 절편과 나)선택적인 마-커(marker)를 코드하는 DNA 서열을 함유하고 있는 적어도 DNA 절편.
  2. 제1항에 있어서, 숙주세포로서 바실러스, 바람직하게는 바실러스 알칼로피러스(Bacillus alcalophilus) 또는 바실러스 리퀘니퍼미스(Bacillus licheniformis)를 사용하는 것을 특징으로 하는 방법.
  3. 제1항에 있어서, 해당 폴리펩타이드를 코드하는 절편 가)의 DNA서열은 프로테아제, 바람직하게는 알카리성 프로테아제를 코드하는 것을 특징으로 하는 방법.
  4. 제1항에 있어서, 해당 폴리펩타이드를 코드하는 절편 가)의 DNA서열은 아밀라제, 바람직하게는 알파-아밀라제를 코드하는 것을 특징으로 하는 방법.
  5. 제1항에 있어서, 항생제 내성을 코드하는 절편 나)에서 선택성 마-커로서 DNA서열이 사용되는 것을 특징으로 하는 방법.
  6. 제5항에 있어서, 항생제인 프레오마이신에 대한 내성을 코드하는 절편 나)에서 선택성 마-커로서 DNA서열이 사용되는 것을 특징으로 하는 방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019930007771A 1992-05-16 1993-05-06 안정성있는 크로모좀 유전자 증폭을 위한 간단한 방법 KR930023461A (ko)

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DE4216246A DE4216246A1 (de) 1992-05-16 1992-05-16 Einfaches Verfahren zur stabilen chromosomalen Genamplifikation
DEP4216246.7 1992-05-16

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KR930023461A true KR930023461A (ko) 1993-12-18

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KR1019930007771A KR930023461A (ko) 1992-05-16 1993-05-06 안정성있는 크로모좀 유전자 증폭을 위한 간단한 방법

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EP (1) EP0570787A3 (ko)
JP (1) JPH06303986A (ko)
KR (1) KR930023461A (ko)
BR (1) BR9301900A (ko)
CA (1) CA2096284A1 (ko)
DE (1) DE4216246A1 (ko)
FI (1) FI932215A (ko)
MX (1) MX9302822A (ko)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU7908998A (en) * 1997-07-03 1999-01-25 Novo Nordisk A/S Protein producing cell containing multiple copies of a desired gene and a screenable marker but no selection marker

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NZ208806A (en) * 1983-07-06 1988-07-28 Gist Brocades Nv Genetic engineering of industrial microorganism species: readily-transformable host used as intermediate in transfer of dna to the industrial species; plasmids
FR2563533B1 (fr) * 1984-04-27 1986-08-22 Centre Nat Rech Scient Procede d'amplification de l'expression d'un gene determine chez bacillus subtilis et souches obtenues
FR2569723B1 (fr) * 1984-08-31 1986-09-19 Centre Nat Rech Scient Application des antibiotiques de la famille des phleomycines a titre d'agent de selection dans le domaine du genie genetique
BR8805646A (pt) * 1987-02-27 1989-10-17 Gist Brocades Nv Processo para amplificacao de genes estaveis em adn cromossomico de microorganismos procarioticos

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JPH06303986A (ja) 1994-11-01
EP0570787A2 (de) 1993-11-24
DE4216246A1 (de) 1993-11-18
BR9301900A (pt) 1993-11-23
CA2096284A1 (en) 1993-11-17
FI932215A (fi) 1993-11-17
EP0570787A3 (en) 1994-08-17
MX9302822A (es) 1993-11-01
FI932215A0 (fi) 1993-05-14

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