Chemistry: analytical and immunological testing – Heterocyclic carbon compound – Hetero-o
Reexamination Certificate
1999-08-30
2001-05-08
Sisson, Bradley L. (Department: 1655)
Chemistry: analytical and immunological testing
Heterocyclic carbon compound
Hetero-o
C436S800000, C435S006120, C435S091100, C536S023100, C536S024300, C536S025300
Reexamination Certificate
active
06228656
ABSTRACT:
The present invention relates to a method of sequence-specifically cleaving nucleic acids as the gene-constituting substance, and is applicable in the fields of clinical diagnosis, cloning of useful genes and exploration of unknown genes.
It is common to cleave nucleic acids at specific sequences in molecular biology and its applications. For example, when a vector carrying a gene encoding a desired protein is constructed for production of the desired protein in microorganisms such as
Escherichia coli
, human cells or other animal cells, it is necessary to cleave a nucleic acid as the gene at a specific sequence with a restriction enzyme.
For such sequence-specific cleavage of nucleic acids, restriction enzymes, which recognize base sequences in nucleic acids and cleave inter-nucleotide linkages, are commonly used, and hundreds of restriction enzymes have already been known.
When restriction enzymes are used, it is necessary to select a restriction enzyme which recognizes a specific sequence to cleave. However, even hundreds of known restriction enzymes are not enough to cleave nucleic acids at any sequences. For example, a promising cancer treatment by specific cleavage of a target cancer gene with the aim of hindering the development of the cancer gene requires a restriction enzyme which specifically cleaves a specific sequence in the cancer gene to be cleaved to attain the aim, but it is possible that there is no restriction enzyme that recognizes and cleaves the specific sequence.
Accordingly, the object of the present invention is to provide a method of specifically cleaving a specific nucleic acid sequence in a double-stranded DNA (hereinafter referred to as a target nucleic acid) which enables cleavage of a specific nucleic acid sequence without using a restriction enzyme.
In order to achieve the above-mentioned object, the present invention provides a method of specifically cleaving a double-stranded DNA (a target nucleic acid) at a specific nucleic acid sequence, which comprises irradiating a solution containing at least the target nucleic acid, a nucleic acid probe (a single-stranded oligonucleotide) linked to an intercalater and spermine with light of an absorption wavelength of the intercalater.
REFERENCES:
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patent: WO88/04301 (1988-06-01), None
patent: WO96/40253 (1996-12-01), None
Ishiguro et al : Nucleic Acids Research, GB Oxford University Press, Surrey vol. 24, No. 24, 1996, pp. 4992-4997, “Fluorescence detection of specific sequence of nucleic acids by Homogenous . . .”.
Le Doan et al Antisene Research and Development vol. 1, No. 1, 1991, pp. 43-54 XP000857736Recognition and photo-induced cleavage and cross-linking of nucleic acids by oligonucleotides etc.
Akerman et al Nucleic Acids Research vol. 24, No. 6, 1996, pp. 1080-1090 XP002125717 Single and double stranded photocleavage of DNA by YO, YOYO and TOTO.
Inoue et al Bioorganic and Medicinal Chemistry (Jun. 1999) 7 (6) 1207-11 XP002125718 Flourescence property of oxazole yellow-linked oligonucleotide etc.
Inoue Teruhiko
Ishiguro Takahiko
Otsuka Masami
Saitoh Juichi
Sugiura Yukio
Nixon & Vanderhye PC
Sisson Bradley L.
Tosoh Corporation
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