KR100956328B1 - Hcv ns5b 의존적인 앱타자임 및 이를 이용한hcv의 진단 방법 - Google Patents
Hcv ns5b 의존적인 앱타자임 및 이를 이용한hcv의 진단 방법 Download PDFInfo
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- KR100956328B1 KR100956328B1 KR1020080027525A KR20080027525A KR100956328B1 KR 100956328 B1 KR100956328 B1 KR 100956328B1 KR 1020080027525 A KR1020080027525 A KR 1020080027525A KR 20080027525 A KR20080027525 A KR 20080027525A KR 100956328 B1 KR100956328 B1 KR 100956328B1
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- C12Q1/701—Specific hybridization probes
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- C12Q1/707—Specific hybridization probes for hepatitis non-A, non-B Hepatitis, excluding hepatitis D
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Abstract
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Claims (8)
- 서열번호 1 내지 4로 기재된 서열 중 1종 이상의 RNA 서열을 포함하는, HCV NS5B 의존적인 앱타자임.
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- HCV NS5B 의존적인 앱타자임의 제조 방법으로서,(a) 교류 모듈에 5개의 랜덤 서열을 포함하는 서열번호 5 및 6의 프라이머 세트(5' 프라이머: 5'-TTGGTGTCTGATGAGNNNNNCGCGCC ATATTGTGAGGGGCGCG-3' 및 3' 프라이머: 5'-ACGTTTGGTTTCCCAAACGTTTCGNN NNNCGCGCCCCTCACAAT-3', N은 G, A, T, C가 동수로 포함됨) 및 taq polymerase로 PCR을 수행하는 단계;(b) 단계 (a)의 PCR 산물에, 서열번호 7 및 8의 프라이머 세트(5'-GGTAATACG ACTCACTATAGGGTTGGTGTCTGATGAG-3' 및 5'-TTTTTTTTTTTTTTTTG GTGTTACGTTTGGTTTCCCAAA-3')와 taq polymerase을 첨가하여, PCR을 수행하는 단계;(c) 단계 (b)의 PCR 산물을 T7 중합효소를 이용하여 시험관내에서 RNA로 전사하여, HCV NS5B 의존적인 앱타자임 라이브러리를 제작하는 단계;(d) 상기 HCV NS5B 의존적인 앱타자임 라이브러리를 NS5B 단백질 이외의 단백질 및 NS5B 단백질과 접촉시켜, NS5B 단백질 이외의 단백질 존재하에서는 특이적인 자가-절단을 보이지 않지만 NS5B 단백질 존재하에서는 특이적 자가-절단을 보이는 RNA 풀을 수득하는 단계;(e) 수득한 앱타자임 RNA 풀 각각을 RT-PCR하여 DNA를 수득하고, 각 염기서열을 확인하는 단계; 및(f) 염기서열을 동정한 각 앱타자임 RNA를 표지한 다음, NS5B 단백질 이외의 단백질 및 NS5B 단백질과 접촉시켜, NS5B 단백질과의 존재하에서만 특이적 자가-절단을 보이는 앱타자임을 선별하는 단계를 포함하는 것을 특징으로 하는 HCV NS5B 의존적인 앱타자임의 제조 방법.
- 제4항에 있어서, 상기 HCV NS5B 의존적인 앱타자임은 서열번호 1 내지 4로 기재된 서열 중 1종 이상의 서열을 포함하는 RNA 서열인 것을 특징으로 하는 HCV NS5B 의존적인 앱타자임의 제조 방법.
- 제1항에 따른 HCV NS5B 의존적인 앱타자임을 포함하는 HCV 진단용 조성물.
- 제6항에 있어서, 상기 조성물은 Mg2+를 더욱 포함하는 것을 특징으로 하는 조성물.
- Mg2+의 존재하에, 제1항에 따른 HCV NS5B 의존적인 앱타자임을 포함하는 HCV 진단용 조성물과 HCV 감염 의심 샘플을 접촉시키는 단계; 및HCV NS5B 의존적인 앱타자임의 자가-절단 반응을 확인하는 단계를 포함하며,HCV NS5B 의존적인 앱타자임의 자가-절단 반응이 확인되면, 상기 샘플을 HCV 양성으로 진단하는 것을 특징으로 하는, HCV 진단을 위한 정보를 제공하는 HCV 검출방법.
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KR1020080027525A KR100956328B1 (ko) | 2008-03-25 | 2008-03-25 | Hcv ns5b 의존적인 앱타자임 및 이를 이용한hcv의 진단 방법 |
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KR100956328B1 true KR100956328B1 (ko) | 2010-05-10 |
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Non-Patent Citations (3)
Title |
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Arch. Biochem. Biophys., Vol.377:1, pp.129~134 (2000) |
J. Virol., Vol.76:8, pp.3688~3696 (2002) |
Nucleic Acid Res., Vol.30:21, pp.4599~4606 (2002) |
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