Optically active DNA probe having phosphonic diester linkage
AssigneeTosch Corporation
InventorsRyuichi Horie, Takahiko Ishiguro
Life of this patent
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Abstract
A DNA probe of the following structural formula (1) (wherein P* is an optically active phosphorus atom, each of R1 and R2 is a DNA oligomer having an arbitrary sequence, and R3 is a fluorescent intercalative dye attached via an appropriate linker) which has an optically active configuration about P*.
Claims12
An optically active DNA probe of formula (1),
- wherein P* is an optically active phosphorus atom, each of R1 and R2 is a DNA oligomer having a desired nucleotide sequence, and R3 is a fluorescent intercalative dye attached via a linker, wherein said probe has an optically active configuration about P*.
An optically active DNA oligomer of formula (2),
- wherein P* is an optically active phosphorus atom, each of R4 and R5 is a DNA oligomer having a desired nucleotide sequence, and R6 is a hydrogen atom or an amino-protecting group, and wherein said probe has an optically active configuration about P*.
A method of preparing an optically active DNA comprising attaching a fluorescent intercalative dye to the nitrogen atom in the aminoethylphosphonate of the optically active DNA oligomer of formula (2) via a linker,
- wherein P* is an optically active phosphorus atom, each of R4 and R5 is a DNA oligomer having a desired nucleotide sequence, and R6 is a hydrogen atom or an amino-protecting group, having an optically active configuration about P*.
A method of preparing an optically active DNA oligomer comprising the steps of:
- (A) reacting an optically active dinucleotide of formula (4),
- wherein P* is an optically active phosphorus atom, B1 and B2 are desired nucleotide bases, R10 is an amino-protecting group, R11 is a hydroxyl-protecting group, and R12 is a hydroxyl-protecting group, a hydrogen atom, a phosphate group or a phosphoroamidite group, with the 5′-hydroxyl group of a first DNA oligomer having a desired nucleotide sequence;
- and then
- (B) reacting a second DNA oligomer having a desired nucleotide sequence to the 5′-terminal.
A method of preparing an optically active dinucleotide of formula (8),
- wherein P* is an optically active phosphorus atom, B1 and B2 are desired nucleotide bases, R16 is an amino-protecting group, R17 is a hydroxyl-protecting group, and R18 is a t-butyldimethylsilyl group, which comprises the steps of:
- (A) reacting a phosphonic nucleotide of formula (5),
- wherein B1 is a desired nucleotide base, R13 is an amino-protecting group, and R14 is a hydroxyl-protecting group, with a nucleoside of formula (6),
- wherein B2 is a desired nucleotide base, and R15 is a t-butyldimethylsilyl group, to give a dinucleotide of formula (7),
- wherein each of B1 and B2 is a desired nucleotide base, R16 is an amino-protecting group, R17 is a hydroxyl-protecting group, and R18 is a t-butyldimethylsilyl group;
- and
- (B) optically resolving the resulting dinucleotide.
Dependencies are inferred from the claim text.