Modified nucleotides and polynucleotides and complexes form therefrom
DC CAFCFirst Claim
1. A compound useful as a probe for detecting the presence or absence of a nucleic acid, said compound having the structure:
- ##STR26## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N1 position of the pyrimidine;
wherein A represents at least three carbon atoms and an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product, and radioisotopes;
wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with detection of A;
wherein if B is a purine, A is attached to the 8-position of the purine, if B is a 7-deazapurine, A is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A is attached to the 5-position of the pyrimidine; and
wherein each of x, y and z represents;
##STR27##
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Abstract
Compounds having the structure: ##STR1## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C1'"'"' -position of the sugar moiety, provided that when B is purine or 7-deazapurine, it is attached at the N9 -position of the purine or 7-deazapurine and when B is pyrimidine, it is attached at the N1 -position;
wherein A represents a moiety consisting of at least three carbon atoms which is capable of forming a detectable complex with a polypeptide when the compound is incorporated into a double-stranded ribonucleic acid, deoxyribonucleic acid duplex, or DNA-RNA hybrid;
wherein the dotted line represents a chemical linkage joining B and A, provided that if B is purine, the linkage is attached to the 8-position of the purine, if B is 7-deazapurine, the linkage is attached to the 7-position of the deazapurine, and if B is pyrimidine, the linkage is attached to the 5-position of the pyrimidine; and
wherein each of x, y and z represents ##STR2## either directly, or when incorporated into oligo- and polynucleotides, provide probes which are widely useful.
Applications include detection and localization of polynucleotide sequences in chromosomes, fixed cells, tissue sections, and cell extracts. Specific applications include chromosomal karyotyping, clinical diagnosis of nucleic acid-containing etiological agents, e.g. bacteria, viruses, or fungi, and diagnosis of genetic disorders.
295 Citations
19 Claims
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1. A compound useful as a probe for detecting the presence or absence of a nucleic acid, said compound having the structure:
- ##STR26## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N1 position of the pyrimidine;
wherein A represents at least three carbon atoms and an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product, and radioisotopes; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with detection of A; wherein if B is a purine, A is attached to the 8-position of the purine, if B is a 7-deazapurine, A is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A is attached to the 5-position of the pyrimidine; and wherein each of x, y and z represents;
##STR27##
- ##STR26## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N1 position of the pyrimidine;
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2. A compound useful as a probe for detecting the presence or absence of a nucleic acid, said compound containing a nucleotide having the structure:
- ##STR28## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that whenever B is purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and whenever B is a pyrimidine, the sugar moiety is attached at the N1 -position of the pyrimidine;
wherein A represents at least three carbon atoms and an indicator molecule selected from the group consisting of fluorescent dyes, electron-dense reagents, enzymes which can be reacted with a substrate to produce a visually detectable reaction product and radioisotopes; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with detection of A; wherein if B is a purine, A or the linkage group is attached to the 8-position of the purine, if B is a 7-deazapurine, A or the linkage group is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A or the linkage group is attached to the 5-position of the pyrimidine; wherein one of x and y represents ##STR29## and the other of x and y is absent or represents --OH or --H; and
wherein z represents H-- or HO--.
- ##STR28## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that whenever B is purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and whenever B is a pyrimidine, the sugar moiety is attached at the N1 -position of the pyrimidine;
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3. A complex useful as a probe for detecting the presence or absence of a nucleic acid, said complex comprising a detectable polypeptide complexed with a compound having the structure:
- ##STR30## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N1 position of the pyrimidine;
wherein A represents at least three carbon atoms, is capable of specifically complexing with the detectable polypeptide when A is linked to B, and represents a component of a signalling moiety capable of producing a detectable signal; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with the characteristic ability of A to form said complex with the detectable polypeptide; wherein if B is a purine, A is attached to the 8-position of the purine, if B is a 7-deazapurine, A is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A is attached to the 5-position of the pyrimidine; and wherein each of x, y and z represents;
##STR31## - View Dependent Claims (4, 5, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
- ##STR30## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety suitable for incorporation into a polynucleotide and covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that when B is a purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and when B is pyrimidine, the sugar moiety is attached at the N1 position of the pyrimidine;
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6. A complex useful as a probe for detecting the presence or absence of a nucleic acid, said complex comprising a detectable polypeptide complexed with an oligo- or polynucleotide containing a nucleotide having the structure:
- ##STR32## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that whenever B is purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and whenever B is a pyrimidine, the sugar moiety is attached at the N1 -position of the pyrimidine;
wherein A represents at least three carbon atoms, is capable of specifically complexing with the detectable polypeptide when A is linked to B, and represents a component of a signalling moiety capable of producing a detectable signal; wherein B and A are covalently attached directly or through a linkage group, said linkage group not interfering substantially with the characteristic ability of A to form said complex with the detectable polypeptide; wherein if B is a purine, A or the linkage group is attached to the 8-position of the purine, if B is a 7-deazapurine, A or the linkage group is attached to the 7-position of the deazapurine, and if B is a pyrimidine, A or the linkage group is attached to the 5-position of the pyrimidine; wherein one of x and y represents ##STR33## and the other of x and y is absent or represents --OH or --H; and
wherein z represents H-- or HO--. - View Dependent Claims (7, 8)
- ##STR32## wherein B represents a purine, 7-deazapurine, or pyrimidine moiety covalently bonded to the C1 '"'"'-position of the sugar moiety, provided that whenever B is purine or 7-deazapurine, the sugar moiety is attached at the N9 -position of the purine or deazapurine, and whenever B is a pyrimidine, the sugar moiety is attached at the N1 -position of the pyrimidine;
Specification