( 2342 of 2347 ) |
United States Patent | 4,617,261 |
Sheldon, III , et al. | October 14, 1986 |
Nucleic acids may be labeled by intercalating the alkylating intercalation moiety of a labeling reagent into a partially double-stranded nucleic acid to form a complex and activating the complex to cause covalent bonding between the reagent and the nucleic acid. Preferably, the labeled nucleic acid is a hybridization probe for detecting nucleic acid sequences capable of hybridizing with a hybridizing region of the nucleic acid. Also preferably the label moiety is non-radioactive. The labeling reagent is of the formula: where A is an alkylating intercalation moiety, B is a divalent organic moiety of the formula: ##STR1## where Y is O, NH or N--CHO, x is a number from 1 to 4, y is a number from 2 to 4, and L is a monovalent label moiety, wherein B is exclusive of any portion of the intercalation and label moieties. Preferably A is a 4-methylene-substituted psoralen moiety, and most preferably A is a 4'-methylene-substituted-4,5',8-trimethylpsoralen moeity and L is biotin.
Inventors: | Sheldon, III; Edward L. (Oakland, CA), Levenson; Corey H. (Oakland, CA), Mullis; Kary B. (Kensington, CA), Rapoport; Henry (Berkeley, CA), Watson; Robert M. (Berkeley, CA) |
Assignee: |
Cetus Corporation
(Emeryville,
CA)
|
Appl. No.: | 06/791,332 |
Filed: | October 25, 1985 |
Application Number | Filing Date | Patent Number | Issue Date | ||
683263 | Dec., 1984 | ||||
591811 | Mar., 1984 | ||||
Current U.S. Class: | 435/6 ; 435/7.24; 435/7.5; 435/7.9; 435/91.1; 436/501; 436/94; 536/24.3; 536/24.31; 536/25.32; 536/25.4; 536/28.5; 536/28.54; 548/303.1; 930/220 |
Current International Class: | C07D 513/04 (20060101); C12Q 1/68 (20060101); C07D 519/00 (20060101); C07H 21/00 (20060101); C07D 513/00 (20060101); C07D 493/00 (20060101); C07D 493/04 (20060101); C12Q 001/68 (); G01N 033/48 () |
Field of Search: | 435/6,7 436/501 935/78 548/303 |
4124598 | November 1978 | Hearst et al. |
4358535 | November 1982 | Falkow et al. |
4542102 | September 1985 | Dattagupta et al. |
0063879 | Nov., 1982 | EP | |||
0131830 | Jan., 1985 | EP | |||
8502628 | Jun., 1985 | WO | |||
2019408 | Oct., 1979 | GB | |||
Sood et al., PNAS, 78, 616-620 (1981). . Ploegh et al., PNAS, 77, 6081-6085. . Brown et al., Gene, 20, 139-144 (1982). . Manning et al., Chromosoma, 53, 107 (1975). . Langer et al., Proc. Natl. Acad. Sci. USA, 78, 6633-6637 (1981). . Leary et al., Proc. Natl. Acad. Sci. USA, 80, 4045-4049 (1982). . Song et al., ANYAS, 355-357 (1980). . Hyde et al., Biochemistry, 17 1251-1257 (1978). . Saffran et al., Proc. Natl. Acad. Sci. USA, 79, 4594-4598 (1982). . Sinha et al., Life Sciences, 17, 1829-1836 (1975). . Hong et al., Cancer Research, 36, 1159-1171 (1976). . Sinha et al., J. Med. Chem., 19, 994-998 (1976). . Cantor, ANYAS, 379-85 (1980). . Schwartz et al., CSH Laboratory Symposium XLVII.. |