KR950011619A - 알레르기질환검출용 dna프로브, 및 그것을 사용한 알레르기 질환유전자의 검출방법 - Google Patents

알레르기질환검출용 dna프로브, 및 그것을 사용한 알레르기 질환유전자의 검출방법 Download PDF

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KR950011619A
KR950011619A KR1019940026915A KR19940026915A KR950011619A KR 950011619 A KR950011619 A KR 950011619A KR 1019940026915 A KR1019940026915 A KR 1019940026915A KR 19940026915 A KR19940026915 A KR 19940026915A KR 950011619 A KR950011619 A KR 950011619A
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미사코 오소에가와
유키 미야베
모토미 나카타
치세이 라
카츠히로 스즈키
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쿠라우찌 노리타카
스미도모덴기고오교오 가부시기가이샤
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Abstract

본 발명에 의하면, 하기 식(1)~(5)로 표시되는 염기배열 및 이들 염기배열과 상보적 염기배열로 이루어진 군으로부터 선택되는 어느 1개의 염기배열을 포함하고, 전체길이 10~50염기의 염기배열로 이루어진 알레르기질환검출용 DNA프로브가 제공된다.
(1) 5' TCC TGC TCT 3'
(2) 5' CTC TCA CCA 3'
(3) 5' ATA TAG GCC 3'
(4) 5' TAG AAT TTC 3'
(5) 5' TGT GAG TGA 3'
또, 본 발명에 의하면, 상기 DNA프로브를 사용해서, 애피니티 크로마토그래피의 수법에 의해 알레르기 질환유전자를 검출하는 방법이 제공된다.

Description

알레르기질환검출용 DNA프로브, 및 그것을 사용한 알레르기 질환유전자의 검출방법
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음

Claims (12)

  1. 하기 식(1)~(5)로 표시되는 염기배열 및 이들 염기배열과 상보적 염기배열로 이루어진 군으로부터 선택되는 어느 1개의 염기배열을 포함하고, 전체길이 10~50염기의 염기배열로 이루어진 것을 특징으로 하는 알레르기질환검출용 DNA프로브.
    (1) 5' TCC TGC TCT 3'
    (2) 5' CTC TCA CCA 3'
    (3) 5' ATA TAG GCC 3'
    (4) 5' TAC AAT TTC 3'
    (5) 5' TGT GAG TGA 3'
  2. 제1항에 있어서, 사람의 고친화성 IgE수용체 β사슬 유전자의 염기배열(1개 사슬 DNA)의 일부와 동일 또는 상보적인 염기배열인 것을 특징으로 하는 알레르기질환검출용 DNA프로브.
  3. 제1항에 있어서, 전체길이 10~50염기의 염기배열의 말단에 표식물질이 부가되어 있는 것을 특징으로 하는 알레르기질환검출용 DNA프로브.
  4. 제1항에 있어서, 전체길이 10~50염기의 염기배열이 말단이, 관능기를 개재해서 불용성 지지체에 결합해서 고정화되어 있는 것을 특징으로 하는 알레르기질환검출용 DNA프로브.
  5. 제1항에 있어서, 전체길이 10~50염기의 염기배열이, 하기 식(6)~(10)의 염기배열 중에서 어느 1개인 것을 특징으로 하는 알레르기질환검출용 DNA프로브.
    (6) 5' CCC AGG AAC TCC TGC TCT TTT TTC A 3'
    (7) 5' GGC TTC CTC TCA CCA GAT ATG TTA 3'
    (8) 5' TGG ATG TGG ATA TAG GCC AAG CTC 3'
    (9) 5' CAT CAT CTC TAC AAT TTC AGT GGA 3'
    (10) 5' TCC ACA GAT TGT GAG TGA CAC AGC A 3'
  6. (a) IgE수용체 유전자를 함유한 피검출 DNA단편을 변성시켜서 1개 사슬로 한후, (b) IgE수용체 유전자의 특정의 영역에 특이적으로 결합하는 DNA프로브와 어니일시켜서, 목적 DNA단편과 DNA프로브와의 결합체를 형성시키고, (c) 그후, 목적 DNA단편과 DNA프로브와의 결합체를 목적으로 하는 DNA단편과, 특이적으로 결합하는 배위자를 고정한 불용성지지체를 고정상으로 하는 컬럼속에서, 상기 불용성지지체와 접촉시켜서, 상기 결합체를 배위자에 결합시키고, (d) 이어서, 컬럼온도를 서서히 상승시키므로서, DNA프로브와 목적 DNA단편과의 결합력을 약화시켜, DNA프로브를 컬럼으로부터 용출시키고, (e) 그 용출패턴을 해석하는 것을 특징으로 하는 알레르기질환유전자의 검출방법.
    단, DNA프로브는, 하기 식(1)~(5)로 표시되는 염기배열 및 이들 염기배열과 상보적 염기배열로 이루어진 군으로부터 선택되는 어느 1개의 염기배열을 포함하고, 전체길이 10~50염기의 염기배열로 이루어진 것이다.
    (1) 5' TCC TGC TCT 3'
    (2) 5' CTC TCA CCA 3'
    (3) 5' ATA TAG GCC 3'
    (4) 5' TAC AAT TTC 3'
    (5) 5' TGT GAG TGA 3'
  7. 제6항에 있어서, IgE수용체 유전자를 함유한 피검출 DNA단편으로서, 사람의 고친화성 IgE수용체 β사슬 유전자를 함유한 피검출 DNA단편을 사용하는 것을 특징으로 하는 알레르기질환유전자의 검출방법.
  8. 제6항에 있어서, IgE수용체 유전자를 함유한 피검출 DNA단편으로서, 건강정상인 또는 알레르기환자의 사람말초형 세포로부터 mRNA를 추출후, cDNA로 변환하고, 이어서, 이 cDNA를 IgE수용체 β사슬을 증폭할 수 있는 프로브를 사용해서 PCR법에 의해서 조제한 cDNA단편을 사용하는 것을 특징으로 하는 알레르기질환유전자의 검출방법.
  9. 제6항에 있어서, 목적으로 하는 DNA단편과 특이적으로 결합하는 배위자로서, 사용하는 DNA프로브의 배열과는 별도의 IgE수용체 β사슬의 유전자 배열 또는 그 상보적 배열로 이루어진 전체길이 10~100 염기의 합성 DNA를 사용하는 것을 특징으로 하는 알레르기질환유전자의 검출방법.
  10. 제9항에 있어서, 배위자로서, 하기의 염기배열 또는 그 상보적 염기배열을 가진 합성 DNA를 사용하는 것을 특징으로 하는 알레르기질환유전자의 검출방법.
    5' CT TCA GGC AGA CTA TTG AAG TCG GCC TCA TCC CCA CCA CTG CAT ACA TGG 3'
  11. 제6항에 있어서, 불용성지지체로서, 트레실클로라이드활성화 실리카겔을 사용하는 것을 특징으로 하는 알레르기질환유전자의 검출방법.
  12. 제6항에 이어서, DNA프로브의 용출패턴으로부터 그 해리온도를 판독하고, 정상 IgE수용체 β사슬 유전자를 함유한 피검출 DNA단편을 사용한 경우와 다른 해리온도를 표시하므로, 피검출 DNA단편이 알레르기질환유전자를 함유한다고 인정하는 것을 특징으로 하는 알레르기유전자의 검출방법.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019940026915A 1993-10-22 1994-10-21 알레르기질환검출용 dna프로브, 및 그것을 사용한 알레르기 질환유전자의 검출방법 KR950011619A (ko)

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