发明公开
EP0530998A1 Detection of complementary nucleotide sequences
失效
Nachweis vonkomplementärenNukleotid-Sequenzen。
- 专利标题： Detection of complementary nucleotide sequences
- 专利标题（中）： Nachweis vonkomplementärenNukleotid-Sequenzen。
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申请号： EP92307441.3申请日： 1992-08-13
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公开(公告)号： EP0530998A1公开(公告)日： 1993-03-10
- 发明人: Eisenbeis, Scott J.
- 申请人： MICROGENICS CORPORATION
- 申请人地址： 2380A Bisso Lane Concord California 94520 US
- 专利权人： MICROGENICS CORPORATION
- 当前专利权人： MICROGENICS CORPORATION
- 当前专利权人地址： 2380A Bisso Lane Concord California 94520 US
- 代理机构： Harrison, David Christopher
- 优先权： US745153 19910815
- 主分类号： C12Q1/68
- IPC分类号： C12Q1/68 ; C12Q1/70 ; G01N33/535
摘要：
The invention relates to a method for detection of a specific nucleic acid sequence which comprises
forming a reaction mixture by combining
(1) a sample suspected of containing a nucleic acid;
(2) a probe/enzyme donor polypeptide conjugate comprising
(a) an enzyme donor polypeptide sequence comprising a β-galactosidase fragment; and
(b) a single-stranded oligonucleotide sequence attached to (a) and capable of hybridizing with said nucleic acid;
(3) an enzyme acceptor polypeptide capable of forming an active enzyme upon complementation with said enzyme donor fragment; and
(4) a substrate for β-galactosidase; and
detecting hybridization of said probe/enzyme donor conjugate to said sample nucleic acid to form a double strand-specific sequence by determining the amount or rate of enzyme activity on said substrate in said reaction mixture. The method can also include a "proof reading" function by incubating the hybridized probe with at least one double-strand-specific, sequence-specific restriction endonuclease.
Novel kits for use in carrying out the method are also included.
forming a reaction mixture by combining
(1) a sample suspected of containing a nucleic acid;
(2) a probe/enzyme donor polypeptide conjugate comprising
(a) an enzyme donor polypeptide sequence comprising a β-galactosidase fragment; and
(b) a single-stranded oligonucleotide sequence attached to (a) and capable of hybridizing with said nucleic acid;
(3) an enzyme acceptor polypeptide capable of forming an active enzyme upon complementation with said enzyme donor fragment; and
(4) a substrate for β-galactosidase; and
detecting hybridization of said probe/enzyme donor conjugate to said sample nucleic acid to form a double strand-specific sequence by determining the amount or rate of enzyme activity on said substrate in said reaction mixture. The method can also include a "proof reading" function by incubating the hybridized probe with at least one double-strand-specific, sequence-specific restriction endonuclease.
Novel kits for use in carrying out the method are also included.
公开/授权文献
- EP0530998B1 Detection of complementary nucleotide sequences 公开/授权日：1996-11-13
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