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KR920004572A - 재조합 진드기 알레르겐 - Google Patents

재조합 진드기 알레르겐 Download PDF

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KR920004572A
KR920004572A KR1019910014784A KR910014784A KR920004572A KR 920004572 A KR920004572 A KR 920004572A KR 1019910014784 A KR1019910014784 A KR 1019910014784A KR 910014784 A KR910014784 A KR 910014784A KR 920004572 A KR920004572 A KR 920004572A
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사또루 오까
가즈히사 오노
세이꼬 시게따
다께시 와다
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오시따 도시아끼
후마낄라 가부시끼가이샤
다나까 류소오
히로시마 다이가꾸
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Abstract

내용 없음

Description

재조합 진드기 알레르겐
본 내용은 요부공개 건이므로 전문내용을 수록하지 않았음
제1도는 이, 콜리(E. coli) Y1090을 숙주로하여 진드기 알레르겐 cDNA가 삽입된 人gtll에 의하여 발현된 진드기 알레르겐 융합 단백질을 SDS－PAGE분석후, 니트로 셀룰로오스 막에 블롯팅하고 단백질 염색과 면역 염색을 한 결과를 나타낸 그림이다; 여기서, 레인 1은 분자량 마커(β－갈락토시다 제 : 분자량 약 1300000)를, 레인 2는 이, 콜리 Y1090세포 전단백질을, 레인 3은 토끼 항－진드기 충체 항원 혈청을 사용한 재조합 진드기 알레르겐의 면역염색(화살표)의 결과를 나타낸다,
제2도는 진드기 알레르겐을 코딩하는 cDAN의 아가로오스 전기영동의 결과를 나타낸 그림이다,
제3도는 제조합 플라스미드 pAK1의 구축을 나타낸 그림이다.

Claims (29)

  1. 진드기 충체 유래 유전자의 발현에 의하여 수득가능한 하기 부분 아미노산 배열(배열번호 : 1)을 함유하는 제조합 진드기 알레르겐.
    Phe Val Met Lys Arg Glu Pro Leu Arg Phe Arg Asp Ile Thr Val Glu
    Gly Asn Glu Asn Ala Tyr Ile Lys Asn Gly Lys Leu His Leu Ser Leu
    Met Asp Pro Ser Thr Leu Ser Leu Val Thr Lys Ala Asp Gly Lys Ile
    Asp Met Thr Val Asp Leu Ile Ser Pro Val Thr Lys Arg Ala Ser Leu
    Lys Ile Asp Ser Lys Lys Tyr Asn Leu Phe His Glu Gly Glu Leu Ser
    Ala Ser Ile
  2. 진드기 충체 유래 유전자의 발현에 의하여 수득가능한 하기 아미노산 배열(배열번호 : 2)을 함유하는 재조합 진드기 알레르겐.
    Phe Val Met Lys Arg Glu Pro Leu Arg Phe Arg Asp Ile Thr Val Glu
    Gly Asn Glu Asn Ala Tyr Ile Lys Asn Gly Lys Leu His Leu Ser Leu
    Met Asp Pro Ser Thr Leu Ser Leu Val Thr Lys Ala Asp Gly Lys Ile
    Asp Met Thr Val Asp Leu Ile Ser Pro Val Thr Lys Arg Ala Ser Leu
    Lys Ile Asp Ser Lys Lys Tyr Asn Leu Phe His Glu Gly Glu Leu Ser
    Ala Ser Ile Val Asn Pro Arg Leu Ser Trp His Gln Tyr Thr Lys Arg
    Asp Ser Arg Glu Tyr Lys Ser Asp Val Glu Leu Ser Leu Arg Ser Ser
    Asp Ile Ala Leu Lys Ile Thr Met Pro Asp Tyr Asn Ser Lys Ile His
    Tyr Ser Arg Gln Gly Asp Gln Ile Asn Met Asp Ile Asp Gly Thr Leu
    Ile Glu Gly His Ala Glu Gly Thr Ile Arg Glu Gly Lys Ile His Ile
    Lys Gly Arg Gln Thr Asp Phe Glu Ile Glu Ser Asn Tyr Arg Tyr Glu
    Glu Gly Val Leu Ser Arg Lys Val Pro Ser His Leu Thr Leu Glu Thr
    Pro Arg Val Lys Met Asn Met Lys Tyr Asp Arg Tyr Ala Pro Val Lys
    Val Phe Lys Leu Asp Tyr Asp Gly Ile His Phe Glu Lys HTs Thr Asp
    Ile Glu Tyr Glu Pro Gly Val Arg Tyr Lys Ile Ile Gly Asn Gly Lys
    Leu Lys Asp Asp Gly Arg His Tyr Ser Ile Asp Val Gn Gly Ile Pro
    Arg Lys Ala Phe Asn Leu Asp Ala Asp Leu Met Asp Lhe Lys Leu Lys
    Val Ser Lys Pro Glu Asp Ser Asn Lys Ala Gln Phe Ser Tyr Thr Phe
    Asn Glu Tyr Thr Glu Thr Glu Glu Tyr Glu Phe Asp Pro His Arg Ala
    Tyr Tyr Val Asn Trp Leu Ser Ser Ile Arg Lys Tyr Ile Gln Asn Phe
    Ile Val Glu Asp Asn
  3. 제1항 또는 제2항에 있어서, 유전자가 데르마토파고이데스 화리나에 유래인 재조합 진드기 알레르겐.
  4. 제1항 내지 제3항중 어느 한 항에 있어서, 알레르겐의 분자량이 SDS－PAGE로 측정시 약 40,000인 재조합 진드기 알레르겐.
  5. 제1항 내지 제4항 중 어느 한 항에 있어서, 알레르겐 cDNA의 총 염기 사슬 길이가 아가로오스 전기 영동법으로 측정시 약 1.2kbp이고, 제4도의 제한 효소지도를 갖는 유전자의 발현에 의하여 수득가능한 재조합 진드기 알레르겐.
  6. 제1항 내지 제5항중 어느 한 항에 따른 재조합 진드기 알레르겐이 다른 단백질과 융합되어 있는 융합 재조합 진드기 알레르겐.
  7. 제6항에 있어서, 다른 단백질이 β－갈락토시다제인 융합 재조합 진드기 알레르겐.
  8. 하기 부분 아미노산 배열(배열번호 : 1)을 함유하는 알레르겐 활성 단백질을 코딩하는 진드기 충체 유래 유전자.
    Phe Val Met Lys Arg Glu Pro Leu Arg Phe Arg Asp Ile Thr Val Glu
    Gly Asn Glu Asn Ala Tyr Ile Lys Asn Gly Lys Leu His Leu Ser Leu
    Met Asp Pro Ser Thr Leu Ser Leu Val Thr Lys Ala Asp Gly Lys Ile
    Asp Met Thr Val Asp Leu Ile Ser Pro Val Thr Lys Arg Ala Ser Leu
    Lys Ile Asp Ser Lys Lys Tyr Asn Leu Phe His Glu Gly Glu Leu Ser
    Ala Ser Ile
  9. 하기 아미노산 배열(배열번호 : 2)을 함유하는 알레르겐 활성 단백질을 코딩하는 진드기 충체 유래 유전자.
    Phe Val Met Lys Arg Glu Pro Leu Arg Phe Arg Asp Ile Thr Val Glu
    Gly Asn Glu Asn Ala Tyr Ile Lys Asn Gly Lys Leu His Leu Ser Leu
    Met Asp Pro Ser Thr Leu Ser Leu Val Thr Lys Ala Asp Gly Lys Ile
    Asp Met Thr Val Asp Leu Ile Ser Pro Val Thr Lys Arg Ala Ser Leu
    Lys Ile Asp Ser Lys Lys Tyr Asn Leu Phe His Glu Gly Glu Leu Ser
    Ala Ser Ile Val Asn Pro Arg Leu Ser Trp His Gln Tyr Thr Lys Arg
    Asp Ser Arg Glu Tyr Lys Ser Asp Val Glu Leu Ser Leu Arg ser Ser
    Asp Ile Ala Leu Lys Ile Thr Met Pro asp Tyr Asn Ser Lys Ile His
    Tyr Ser Arg Gln Gly Asp Gln Ile Asn Met Asp Ile Asp Gly Thr Leu
    Ile Glu Gly His Ala Gln Gly Thr Ile Arg Glu Gly Lys Ile His Ile
    Lys Gly Arg Gln Thr Asp Phe Glu Ile Glu Ser Asn Tyr Arg Tyr Glu
    Asp Gly Lys Leu Ile Ile Glu Pro Val Lys Ser Glu Asn Gly Lys Leu
    Glu Gly Val Leu Ser Arg Lys Val Pro Ser His Leu Thr Leu Glu Thr
    Pro Arg Val Lys Met Asn Met Lys Tyr Asp Arg Tyr Ala Pro Val Lys
    Val Phe Lys Leu Asp Tyr Asp Gly Ile His Phe Glu Lys His Thr Asp
    Ile Glu Tyr Glu Pro Gly Val Arg Tyr Lys Ile Ile Gly Asn Gly Lys
    Leu Lys Asp Asp Gly Arg His Tyr Ser Ile Asp Val Gln Gly Ile Pro
    Arg Lys Ala Phe Asn Leu Asp Ala Asp Leu Met Asp Phe Lys Leu Lys
    Val Ser Lys Pro Glu Asp Ser Asn Lys Ala Gln Phe Ser Tyr Thr Phe
    Asn Glu Tyr Thr Glu Thr Glu Glu Tyr Glu Phe Asp Pro His Arg Ala
    Tyr Tyr Val Asn Trp Leu Ser Ser Ile Arg Lys Tyr Ile Gln Asn Phe
    Ile Val Glu Asp Asn
  10. 제8항에 있어서, 유전자가 하기 DNA 배열(배열번호 :1)을 함유하는 진드기 충체 유래 유전자.
    TTT GTC ATG AAA CGA GAA CCA TTG CGA TTC AGA GAC ATC ACT GTC GAA
    GGA AAC GAA AAT GCC TAT ATC AAA AAT GGC AAA CTT CAT TTG TCG CTT
    ATG GAT CCG TCA ACA TTG AGT TTA GTC ACG AAA GCC GAT GGA AAA ATC
    GAC ATG ACA GTA GAC TTG ATA TCG CCA GTC ACA AAA CGT GCA TCG TTG
    AAA ATT GAT TCA AAG AAA TAC AAC CTT TTC CAT GAA GGT GAA TTG AGT
    GCA TCG ATC
  11. 제9항에 있어서, 유전자가 하기 DNA 배열(배열번호 : 2)을 함유하는 진드기 충체 유래 유전자.
    TTT GTC ATG AAA CGA GAA CCA TTG CGA TTC AGA GAC ATC ACT GTC GAA
    GGA AAC GAA AAT GCC TAT ATC AAA AAT GGC AAA CTT CAT TTG TCG CTT
    ATG GAT CCG TCA ACA TTG AGT TTA GTC ACG AAA GCC GAT GGA AAA ATC
    GAC ATG ACA GTA GAC TTG ATA TCG CCA GTC ACA AAA CGT GCA TCG TTG
    AAA ATT GAT TCA AAG AAA TAC AAC CTT TTC CAT GAA GGT GAA TTG AGT
    GCA TCG ATC GTA AAC CCA CGA TTG TCA TGG CAT CAA TAC TCG AAA CGC
    GAT TCT CGT GAA TAC AAG AGT GAT GTA GAA CTA TCG TTG CGA TCG TCG
    GAC ATT GCT CTC AAG ATT ACG ATC CCT GAT TAT AAT TCG AAA ATT CAT
    TAT TCA CGA CAA GGT CAA ATC AAC ATG GAC ATC GAT GGT ACA TTG
    ATC GAA GGT CAT GCA CAA GGA ACC ATC AGA GAA GGT AAA ATC CAC ATT
    AAA GGT AGA CAA ACT GAT TTC GAG ATC GAA TCC AAC TAC CGA TAC GAA
    GAT GGC AAA CTA ATC ATC GAA CCG GTC AAG AGT GAA AAT GGC AAA TTG
    GAA GGC GTT CTT TCC CGT AAG GTG CCA TCA CAT CTG ACA CTA GAA ACA
    CCA CGA GTC AAG ATG AAT ATG AAA TAT GAT CGA TAT GCA CCA GTC AAA
    GTG TTC AAA TTG GAT TAT GAT GGC ATC CAC TTC GAG AAA CAT ACC GAT
    ATT GAA TAC GAA CCT GGC GTT CGA TAC AAG ATC ATC GGC AAT GGA AAA
    CTC AAG GAT GAT GGC CGC CAC TAT TCT ATC GAT GTC CAA GGT ATT CCA
    CGC AAA GCA TTC AAT CTG GAC GCT GAC TTG ATG GAT TTC AAA CTG AAA
    GTG AGC AAG CCA GAA GAT AGC AAT AAA GCT CAA TTC AGC TAC ACA TTC
    AAC GAA TAT ACC GAG ACC GAA GAA TAT GAA TTC GAT CCA CAT CGT GCC
    TAT TAT GTT ATT TGG TTG AGT TCC ATT CGC AAA TAC ATC CAG AAT TTC
    ATC GTC GAA GAC AAC
  12. 제10도로 표시되는 염기 배열에서, 적어도 상류로부터 약 170bp 내지 약 270bp, 약 270bp 내지 약 400bp 또는 약 170bp 내지 약 400bp의 영역내에 코딩되는 아미노산 배열을 함유하는 진드기 알레르겐 단편.
  13. 진드기 충체 유래 진드기 알레르겐에 함유된 에피토우프를 갖는 폴리펩티드.
  14. 진드기 충체 유래 진드기 알레르겐에 함유된 에피토우프와 면역학적으로 동일하다고 간주될 수 있는 에피토우프를 갖는 폴리펩티드.
  15. 제13항 또는 제14항에 있어서, 하기 아미노산 배열(배열번호 : 3)을 함유하는 폴리펩티드.
    Met Thr Val Asp Leu Ile Ser Pro Val Thr Lys Arg Ala Ser Leu Lys
    Ile Asp Ser Lys Lys Tyr
  16. 제13항 또는 제14항에 있어서, 하기 아미노산 배열(배열번호 : 4)을 함유하는 폴리펩티드.
    Asp Val Glu Leu Ser Leu Arg Ser Ser Asp Ile Ala
  17. 제15항 또는 제16항의 폴리펩티드를 코딩하는 유전자.
  18. 제17항에 있어서, 하기 DNA 배열(배열번호 : 3)을 함유하는 유전자.
    ATG ACA GTA GAC TTG ATA TCG CCA GTC ACA AAA CGT GCA TCG TTG
    AAA ATT GAT TCA AAA TAC
  19. 제17항에 있어서, 하기 DNA 배열(배열번호 : 4)를 함유하는 유전자.
    GAT GTA GAA CTA TCG TTG CGA TCG TCG GAC ATT GCT
  20. 형질전환된 세균, 효모 또는 포유동물 세포중에서 제8항 내지 제11항 중 어느 한 항의 유전자를 발현시킬수 있는 발현 벡터.
  21. 제20항의 발현벡터로 형질전환된 세균, 효모 또는 포유동물 세포.
  22. 제21항의 세균, 효모 또는 포유동물 세포를 이 유전자가 발현 가능한 조건하에서 배양하여 재조합 진드기 알레르겐을 생성시키고, 이어서 이 재조합 진드기 알레르겐을 회수함을 특징으로 하는 재조합 진드기 알레르겐의 재조방법.
  23. 제21항의 세균, 효모 또는 포유동물 세포를 이 유전자가 발현가능한 조건하에서 배양하여 재조합 진드기 알레르겐을 생성시키고,이 융합 재조합 진드기 알레르겐을 회수하고, 이어서 융합되어 있는 다른 단백질을 탈리시킴을 특징으로 하는 재조합 진드기 알레르겐의 제조방법.
  24. 제1항 내지 제5항중 어느 한 항의 재조합 진드기 알레르겐을 유효성분으로 하는 진드기 알레르기 질환 치료제.
  25. 제1항 내지 제5항중 어느 한 항의 재조합 진드기 알레르겐을 유효성분으로 하는 진드기 알레르기 질환 진단약.
  26. 제12항의 진드기 알레르겐 단편을 유효성분으로 하는 진드기 알레르기 질환 치료제.
  27. 제12항의 진드기 알레르겐 단편을 유효성분으로 하는 진드기 알레르기 질환 진단약.
  28. 제15항 또는 제16항의 폴리펩티드를 유효성분으로 하는 진드기 알레르기 질환 치료제.
  29. 제15항 또는 제16항의 폴리펩티드를 유효성분으로 하는 진드기 알레르기 질환 진단약.
    ※ 참고사항 : 최초출원 내용에 의하여 공개하는 것임.
KR1019910014784A 1990-08-27 1991-08-26 재조합 진드기 알레르겐 KR0148020B1 (ko)

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JP91-104949 1991-03-09
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EP0473111A3 (en) 1992-09-02
US5314991A (en) 1994-05-24
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AU649704B2 (en) 1994-06-02
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US5405758A (en) 1995-04-11
AU8271091A (en) 1992-03-05

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