Detection of complementary nucleotide sequences
US5795718A · kind A · utility
Assignee
Inventor
Key dates
| Filing date | May 30, 1995 |
| Grant date | Aug 18, 1998 |
| Priority date | — |
| Expiry date | May 30, 2015 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12Q1/6816
- WIPO fieldBiotechnology
- WIPO sectorChemistry
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 PA1 (a) an enzyme donor polypeptide sequence comprising a .beta.-galactosidase fragment; and PA1 (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.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.