Detection of complementary nucleotide sequences
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United States of America Patent
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Aug 18, 1998
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N/A
app pub date -
May 30, 1995
filing date -
Aug 15, 1991
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Abstract
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 .beta.-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 .beta.-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.
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