Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1995-02-09
2000-01-11
Houtteman, Scott W.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 266, 536 284, C12Q 168
Patent
active
060134348
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to novel oligonucleotide-polyamide conjugates preferably having a free 3' hydroxyl moiety, and wherein the polyamide is coupled to the oligonucleotide through its carboxyl terminus.
BACKGROUND ART
A number of molecular biological methods involve the detection of nucleotide sequences, or the tagging of nucleotide sequences with reporter groups. For example, the widely utilised polymerase chain reaction technique (PCR) allows many copies of a desired DNA to be generated from only a few target molecules within a period of 1 to 2 hours and consequently has found applications of nucleic acid detection techniques previously limited by low sensitivity. Detection of amplified PCR products may be carried out in a number of ways. It is most desirable to be able to detect amplification products non-radioactively, for example, using a reporter group within the amplified nucleotide sequences.
Similarly, in reactions such as nick translation, it is desirable to incorporate a labelled nucleotide into reaction products produced by DNA polymerase or RNA polymerase for subsequent detection of hybridisation products.
DISCLOSURE OF INVENTION
In accordance with the present invention there is provided in one aspect a nucleotide polyamide conjugate of the formula (I): alkylene group, in which one or more carbons may optionally be replaced by --NH--, --O-- or --S--, X.sup.2 is a bond, or an unsubstituted or substituted C.sub.1 -C.sub.20 alkylene group, in which one or more carbons may optionally be replaced by --NH--, --O-- or --S--, the optional substituents in X.sup.1 or X2 selected from one or more of: oxo, amino, thioxo, hydroxyl, mercapto, carboxyl, halogen, lower alkyl, phenyl, amino-lower alkyl, ester-lower alkyl, amido-lower alkyl, ether-lower alkyl, or thioether-lower alkyl, groups, the sulfur analogues of these substituents, or the side-chain substituents from naturally occurring amino acids, and the closely related analogues of these sidechains, for example. X.sup.3 is an amino acid, or a polyamide linked via its carboxy terminus, NUC is a nucleoside of any one of the formulas: ##STR1## where .fwdarw.indicates the bond to the --C.tbd.C-- group in formula (I), and X.sup.4 is a sugar group of the formula: ##STR2## where the 5' oxygen is linked to Nu, and X.sup.5 and X.sup.6 are each independently, H or OR, where R is H, a protecting group, or a solid phase matrix, and Nu is an oligonucleotide.
X.sup.1 is an unsubstituted or substituted C.sub.1 -C.sub.10 alkylene group, in which one or more carbons may optionally be replaced by --NH--, --O-- or --S--, and the optional substituents in X.sup.1 are selected from one or more of oxo, amino, thioxo, hydroxyl, mercapto, carboxyl, halogen, lower alkyl, phenyl, amino-lower alkyl, ester-lower alkyl, amido-lower alkyl, ether-lower alkyl, or thioether-lower alkyl, groups, and the like, such as the sulfur analogues of these compounds, and any other functional groups. Other possible substituents are the side-chain substituents from naturally occurring amino acids, and their closely related analogues, for example. Preferably, the C.sub.1 to C.sub.10 alkylene is C.sub.1-3 alkylene and is optionally substituted with one or more of amide, halogen, aryl, ester and the like. A preferred form of X.sup.1 is methylene.
X.sup.2 is a bond, or an unsubstituted or substituted C.sub.1 -C.sub.20 alkylene group, in which one or more carbons may optionally be replaced by --NH--, --O-- or --S--, and the optional substituents in X.sup.2 are selected from one or more of oxo, amino, thioxo, hydroxyl, mercapto, carboxyl, halogen, lower alkyl, phenyl, amino-lower alkyl, ester-lower alkyl, amido-lower alkyl, ether-lower alkyl, or thioether-lower alkyl, groups, the sulfur analogues of these substituents, or the side-chain substituents attached to the .alpha.-carbon of naturally occuring amino acids, and their similar analogues, for example. X.sup.2 preferably has the form --CO--(C.sub.1 to C.sub.9 alkylene)--NH--, where the alkylene may be further substituted, for
REFERENCES:
patent: 5427914 (1995-06-01), Dennis
Haralambidis Jim
Tregear Geoffrey William
Houtteman Scott W.
Howard Florey Institute of Experimental Physiology and Medicine
LandOfFree
Oligonucleotide-polyamide conjugates does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Oligonucleotide-polyamide conjugates, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Oligonucleotide-polyamide conjugates will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1461119