KR910009926A - 연결 가능한 헤어핀 프로브와 전사를 이용한 핵산 증폭 방법 - Google Patents

연결 가능한 헤어핀 프로브와 전사를 이용한 핵산 증폭 방법 Download PDF

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KR910009926A
KR910009926A KR1019900018126A KR900018126A KR910009926A KR 910009926 A KR910009926 A KR 910009926A KR 1019900018126 A KR1019900018126 A KR 1019900018126A KR 900018126 A KR900018126 A KR 900018126A KR 910009926 A KR910009926 A KR 910009926A
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sequence
target sequence
rna polymerase
probe
hairpin
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다타굽타 나니브후산
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고든 피. 폴리
몰레큘라 다이어그노스틱스, 인크.
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    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract

내용 없음

Description

연결 가능한 헤어핀 프로브와 전사를 이용한 핵산 증폭 방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1-3도는 헤어핀 형태 및 다이머 형태를 포함한 본 발명의 여러 상이한 형태의 프로브를 예시한 것이다.

Claims (9)

  1. (1) 혼성화 조건하에서 프로머터 기능 영역을 갖는 헤어핀 구조를 형성할 수 있는 단일 가닥 자기 상보성 서열 및 (2) 자기 상보성 서열의 3′말단으로부터 연장된 단일 가닥으로 된 프로브 서열을 포함하며, 상기 프로브 서열에 상보적인 표적 서열을 혼성화시키고, 생성된 혼성화 표적 서열을 헤어핀 형성 자기 상보성 서열의 5′말단에 연결시킬 때 상기 표적 서열이 헤어핀 형성 혼성화 조건하에서 RNA 폴리머라제 및 필요한 리보뉴클레오시트 트리포스페이트 존재하에 전사가능한 것을 특징으로 하는 핵산 프로브.
  2. 제1항에 있어서, DNA로 구성되고, 상기 폴리머라제가 DNA-의존성 RNA 폴리머라제, 바람직하기로는 T7, T3 또는 SP6 박테리오파지 RNA 폴리머라제이고, 바람직하기로는 상기 프로모터 영역이 아래와 같은 헤어핀 형태를 갖는 염기 서열을 포함하는 것을 특징으로 하는 프로브.
    상기 염기 서열에 있어서 n은 2 내지 50의 정수이다.
  3. (A) 액체 혼합물 중에서 표적 서열을 제1항 기재의 프로브와 혼성화시키는 단계, (B) 생성된 혼성화 표적 서열을 헤어핀 형성 자기 상보성 서열의 5′말단에 연결시키는 단게, (C) 연결된 혼성체에 혼성체에서 표적 서열를 전사시키기에 충분한 RNA 폴리머라제 및 리보뉴클레오시드 트리포스페이트를 첨가하는 단계, (D) 예정된 시간 동안 상기 표적 서열의 전사가 일어나도록 허용함으로써 상기 표적 서열의 증폭물로서 생성된 상보성 RNA 전사 생성물을 모으는 단계로 이루어지는, 특정한 표적 핵산 서열을 증폭시키는 방법.
  4. 제3항에 있어서, 생성된 RAN 전사 생성물을 제2의 증폭 단계에서 추가 증폭시키는 것을 특징으로 하는 방법.
  5. 제3항 내지 4항에 있어서, 상기 폴리머라제가 DNA-의존성RNA 폴리머라제, 바람직하기로는 T7, T3 또는 SP6 박테리오파지 RNA폴리머라제이고; 바람직하기로는 상기 프로모터 영역이 아래와 같은 헤어핀 형태를 갖는 염기 서열을 포함하는 것을 특징으로 하는 방법.
    상기 염기 서열에 있어서, n은 2 내지 50의 정수이다.
  6. (A) 시료중의 표적 서열을 액체 혼합물 중에서 제1항 기재의 프로브와 혼성화시키는 단계, (B) 생성된 혼성화 표적 서열을 헤어핀 형성 자기 상보성 서열의 5′말단에 연결시키는 단계, (C) 연결된 서열에 혼성체내에서 표적 서열를 전사시키기에 충분한 RNA 폴리머라제 및 리보뉴클레오시드 트리포스페이트를 첨가하는 단게, (D) 예정된 시간 동안 표적 서열의 전사가 일어나도록 허용함으로써 생성된 삼보성 RNA 전사 생성물을 모으는 단계, (E) 모은 RNA 전사 생성물을 검출하는 단계로 이루어진 시료 중의 특정 표적 핵산 서열을 검출하는 방법.
  7. 제6항에 있어서, 생성된 RNA 전사 생성물을 제2의 증폭 단계에서 추가 증폭시키는 것을 특징으로 하는 방법.
  8. 제6항 내지 7항에 있어서, 상기 프로모터 영역이 아래와 같은 헤어핀 형태를 갖는 염기 서열을 포함하는 것을 특징으로 하는 방법.
    상기 염기 서열에 있어서, n은 2 내지 50의 정수이다.
  9. (1) 표적 서열과 혼성화시키고 연결시켰을 때, RNA 폴리머라제 및 리보뉴틀레오시드 트리포스페이트의 존재하에서 전사가능한 제1항 또는 2항 기재의 핵산 프로브. (2) 상기 RNA 폴리머라제로 이루어진, 특정 핵산 서열의 증폭에 유용한 시약 키트.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019900018126A 1989-11-09 1990-11-09 연결 가능한 헤어핀 프로브와 전사를 이용한 핵산 증폭 방법 KR910009926A (ko)

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US43437289A 1989-11-09 1989-11-09
US434372 1989-11-09
US07/569,991 US5215899A (en) 1989-11-09 1990-08-23 Nucleic acid amplification employing ligatable hairpin probe and transcription
US569991 1990-08-23

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US (1) US5215899A (ko)
EP (1) EP0427073A3 (ko)
JP (1) JPH048293A (ko)
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AU (1) AU645996B2 (ko)
FI (1) FI905517A0 (ko)
IE (1) IE904029A1 (ko)
IL (1) IL96251A0 (ko)
NO (1) NO904634L (ko)
NZ (1) NZ235980A (ko)
PT (1) PT95817A (ko)

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