WO2001038571A1 - Molecule reconnaissant les mesappariements - Google Patents
Molecule reconnaissant les mesappariements Download PDFInfo
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- WO2001038571A1 WO2001038571A1 PCT/JP2000/007497 JP0007497W WO0138571A1 WO 2001038571 A1 WO2001038571 A1 WO 2001038571A1 JP 0007497 W JP0007497 W JP 0007497W WO 0138571 A1 WO0138571 A1 WO 0138571A1
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- base pair
- pair
- base
- carbon atoms
- normal
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6832—Enhancement of hybridisation reaction
Definitions
- the present invention provides a method for forming a pseudo base pair between a base pair that cannot form a normal base pair and a base pair that cannot form a normal base pair.
- a method for detecting and identifying a base pair that cannot form a normal base pair by measuring base pair formation, a reagent therefor, a kit containing the same, a compound thereof, and a method using the method The present invention relates to a method for detecting an abnormality in the nucleotide sequence of DNA or RNA.
- bases forming pairs are determined.
- guanine (G) has cytosine (C) and adenine (A) has thymine (T).
- G cytosine
- A adenine
- T thymine
- most bases can form such a pair under conditions that allow one DNA to hybridize to another, while one or several bases can form such a pair. May not be able to form a perfect pair.
- Such base pairs that cannot form ordinary base pairs are hereinafter referred to as mismatches in the present specification.
- the method of detecting such a mismatch is generally a method of comparing the hybridization efficiency of double-stranded DNA.
- the use of this method requires a great deal of effort because it is necessary to know the nucleotide sequence of DNA, including Misuchi, in advance, and is unsuitable as a method for treating many samples.
- a DNA repair protein such as MutS
- bulge DNA recognition molecule that specifically binds to and stabilizes DNA (bulge DNA) having an unpaired base (bulge base) generated in double-stranded DNA.
- This bulge recognition molecule not only forms hydrogen bonds with unpaired bases, but also uses the stacking interaction between the aromatic ring and bases near the bulge in the space created by the presence of the bulge base, thereby forming an interface. It is stable and stable.
- the present inventors have further studied the effect on unpaired bases using the stacking effect due to the presence of such bases in the vicinity. It has been found that a compound having two pair-forming molecular species can be relatively stably incorporated by such a stacking effect. Disclosure of the invention
- the present invention provides a method capable of detecting such a base-pair mismatching simply and with high sensitivity.
- the present invention provides a method capable of easily and easily detecting a mismatch existing in a double-stranded DNA chain and a detection reagent therefor.
- the present invention relates to a base pair that cannot form a normal base pair, represented by the following general formula (I): A-L-B (I)
- A is a chemical structural part capable of forming a pair with one of the base pairs that cannot form a normal base pair
- B is a chemical structure part that cannot form a normal base pair
- L a chemical structural part capable of forming a pair with the other base, represents a linker structure linking chemical structural parts A and B.
- the base pair that cannot form a base pair is formed by measuring the formation of the pseudo base pair by causing the base pair that cannot form a base pair to form a pseudo base pair. It concerns detection and identification methods.
- the present invention provides a method for measuring a normal base comprising forming a pseudo-base pair with the above-mentioned base pair which cannot form a normal base pair, and measuring the formation of the pseudo base pair.
- a method for detecting and identifying a base pair that cannot form a pair the following general formula (I) for causing a base pair that cannot form a normal base pair to form a pseudo-base pair: ,
- A is a chemical structural part that can form a pair with one of the base pairs that cannot form a normal base pair
- B is another chemical group that cannot form a normal base pair
- L a chemical structural part capable of forming a pair with one base, represents a linker structure that connects chemical structural parts A and B.
- the present invention relates to a reagent consisting of a compound represented by the formula: which is for forming a pseudo base pair with a base pair which cannot form a normal base pair.
- the present invention provides a method for forming a pseudo base pair on a base pair which cannot form a normal base pair comprising the reagent of the present invention and a material for detection and identification, and forms the pseudo base pair.
- the present invention relates to a kit for detecting and identifying a base pair which cannot form a normal base pair by measuring the formation of base pairs.
- R, R> is a hydrogen atom, an alkyl group having 1 to 15 carbon atoms, and one or more carbon atoms in the alkyl group may be substituted with an oxygen atom or a nitrogen atom.
- R 2 represents an alkyl group having 1 to 20 carbon atoms, wherein one or more carbon atoms in the alkyl group may be substituted with an oxygen atom, a nitrogen atom or a carbonyl group.
- an immobilized substance modified to a chemical structure which can be immobilized on a plate or a detection device for instrumental analysis.
- the present invention hybridizes a single-stranded DNA or RNA as a specimen with a corresponding DNA or RNA having a normal base sequence, and then forms a normal base pair in the hybridized DNA or RNA.
- a base pair that cannot be converted is represented by the following general formula (I),
- A is a chemical structural part that can form a pair with one of the base pairs that cannot form a normal base pair
- B is another chemical group that cannot form a normal base pair
- L a chemical structural part capable of forming a pair with one base, indicates a linker structure that connects chemical structural parts A and B.
- the present invention relates to a method for detecting and identifying a base pair that cannot form a normal base pair by measuring the formation of a basic base pair, and a method for detecting a base sequence abnormality in DNA or RNA.
- FIG. 1 is a photograph instead of a drawing, showing the inhibitory effect of DNase I on cleavage of a mismatch site by a mismatch recognition molecule of the present invention.
- FIG. 2 is a graph showing the inhibitory effect of DNasel on cleavage when the mismatch recognition molecule of the present invention is used.
- FIG. 3 schematically shows the action of the mismatch recognition molecule of the present invention at the mismatch portion.
- the present inventors have developed a pargi DNA recognition molecule that specifically binds and stabilizes DNA (bulge DNA) having an unpaired base (bulge base) generated in double-stranded DNA.
- This bulge-recognition molecule is stabilized by intercalation in the space created by the presence of the bulge base by utilizing the stacking interaction between the aromatic ring and the base near the bulge.
- the present inventors connect two such bulge recognition molecules with a binding chain such as a linker so that each bulge recognition molecule is bulge-recognized at a base-pair mismatching portion.
- This compound pairs with guanine at the 1,8-naphthyridine moiety.
- guanine is a bulge base, there is sufficient space for the guanine to form a pair with the 1,8-naphthyridine derivative.
- the formation of a pair is sufficient if the stability of both is considered, but in the case of a mismatch, there is not enough spatial margin because another base already exists at the place for forming the pair.
- whether or not such a relatively large molecular species can stably enter a small space between a base and an adjacent base is a major problem.
- the DNase I DNA hydrolase footblanting titration in the presence of the compound of the formula (III) at various concentrations was performed to measure the DNA by DNase I. The location of cleavage inhibition was investigated.
- FIG. 1 is a photograph replacing a drawing showing the results of electrophoresis.
- the concentration of the compound of formula (III) gradually increases from 0 to 500 / iM.
- DNasel DNA hydrolase
- Such a DNA cleavage inhibitory effect on DNA hydrolase depends on the presence (including concentration) of the compound of formula (III) and is a specific action of the compound of formula (III). it is conceivable that.
- Figure 2 shows the relationship between the strength of the cutting band and the concentration of added naphthyridine in Figure 1.
- the vertical axis in Fig. 2 is the cutting inhibition ratio obtained from the strength of the cutting band, where "0.0" indicates that the cutting is almost complete, and "1.0" indicates that the cutting is almost complete. This shows the situation where cleavage was inhibited.
- the horizontal axis in Figure 2 is added The concentration (M) of the compound of formula (III).
- the black circles (•) in the graph of Fig. 2 are from the G-G Miss Multisite, and the black triangles ( ⁇ ) are from the G-A Miss Multisite.
- equation binding constant to mismatch GG compound of ( ⁇ ) (K a (GG mis)) is 1.1 3 determined to be x 1 0 7 M, likewise in G-A to mismatch binding constant (K a (GAm is)) were asked 1. and 6 3 xl 0 4 M 1.
- the ratio of the binding constants of the two ((K a (G Gm is)) / (K a (GAm is))) is 696, and the formula (III) shows that the specificity for G—G mismatch You can see that it is working. Moreover, the fact that the binding constant for the G-G mismatched base pairs of the compound of formula (III) is relatively large on the order of 1 0 7, formula (III) stably G-G Misumadzuchi base pairs compound than expected Indicates that it has been incorporated into the part.
- the Mismatsu base-recognizing molecule of the present invention incorporated into the double-stranded DNA forms a relatively stable pair, and the natural enzyme cannot recognize the pair due to the formation of such a pair. It is considered that the base sequence is newly formed.
- FIG. 3 schematically shows that the compound (mismatch recognition molecule) represented by the general formula (I) of the present invention is relatively stably incorporated into the mismatched portion of the base.
- the left side of FIG. 3 shows a portion of the double-stranded DNA having a G—A mismatch. In other places, normal base pairs are formed, and the DNA is hybridized as a whole in spite of mismatches at the G-A portion. To this, NN.
- the mismatch recognition molecule of the present invention is added, the state shown in the right side of FIG. 3 is considered. That is, the bases are mismatches Guanin (G) is Guanin recognition site (N G) and form a pair of mismatched recognition molecule
- the other mismatched base, adenine (A) forms a pair with the adenine recognition site (N A ) of the mismatch recognition molecule, and the guanine recognition site (N.) of the mismatch recognition molecule.
- the recognition site (N A ) is a linker of a suitable length and flexibility with a suitable degree of freedom, and almost any other normal base in the double-stranded DNA strand. It is considered that they are taken in the same way as pairs (see the right side of Fig. 3).
- Another major reason why the mismatch recognition molecule of the present invention is relatively stably incorporated into the double-stranded DNA chain is that the mismatch recognition molecule has a base recognition site (for example, the guanine recognition site ( This means that N 0 ) and the adenine recognition site (NA)) are stabilized by the stacking effect of the preceding and following bases (such as the intermolecular force between bases).
- the base recognition site (the chemical structure portion of A and B in the general formula (I)) of the mismatch recognition molecule of the present invention is not limited to simply forming a hydrogen bond with the target base, but also before, after, or around. It must be a chemical structure that can achieve the stacking effect of different bases.
- a compound in which two base recognition sites are linked by a linker having an appropriate length and an appropriate degree of freedom is specific to a base pair mismatch portion in a double-stranded nucleic acid.
- the basic concept is to find that a pair is formed efficiently and stably, and is not limited to the GG mismatch described above.
- mismatch recognition is not limited to GG mismatch.
- the base recognition site in the mismatch recognition molecule recognizes one of the mismatched bases and recognizes the base and the Watson leak (Wa). (tson-Crick) type base pairs, and by selecting a molecular species capable of obtaining a stacking effect by surrounding bases, various bases and base pairs are not limited to the exemplified guanine. What is necessary is just what can form.
- mismatched base when the mismatched base is cytosine, 2-aminoaminophthyridine-141-one or a derivative thereof is used as the base recognition site, and when the mismatched base is adenine, the 2- quinolone derivative is used. For example, 3- (2-aminoethyl) -12-quinolone or a derivative thereof is used.
- mismatched base is thymine, 2-aminononaphthyridin-17-one or a derivative thereof is used. .
- the base recognition site in the mismatch recognition molecule of the present invention which is specifically recognized by a specific mismatched base, has a hydrogen bonding site for forming a hydrogen bond and a planar structure for being stacked by a nearby base.
- a compound having a heterocyclic aromatic group is preferred, and a heterocyclic group having a substituent having a certain degree of steric hindrance is more preferable in order to enhance selectivity for a base.
- Examples of such a substituent include a linear or branched alkyl group having 1 to 15 carbon atoms, preferably 1 to 10 or more preferably 1 to 7, and 1 to 15 carbon atoms. 15, preferably 1 to 10, more preferably 1 to 7, an alkoxy group comprising a linear or branched alkyl group, having 1 to 15 carbon atoms, preferably 1 to 10 More preferably, a mono- or di-alkylamino group mono- or di-substituted with 1 to 7 linear or branched alkyl groups is exemplified.
- One or more carbon atoms in these alkyl, alkoxy or mono- or dialkylamino groups may be replaced by oxygen or nitrogen atoms.
- the linker part L in the compound represented by the general formula (I) of the present invention is not particularly limited as long as the two base recognition sites have an appropriate length and an appropriate degree of freedom.
- a linear or branched saturated or unsaturated alkyl group having 1 to 20 carbon atoms, preferably 1 to 15 carbon atoms, and more preferably 1 to 12 carbon atoms.
- a preferred linker is one having an amide bond at both ends as in the compound of formula (III) described above, and a nitrogen at the center. And those having an elementary atom.
- This linker part can bind not only two base recognition sites, but also a branch for immobilization to a carrier from the linker part.
- a branch such as an alkylene group having a functional group or the like for binding to the carrier from the position of the nitrogen atom near the center of the linker, and further immobilize the branch to the carrier if necessary. it can.
- the bond between the base recognition site A or B and the linker part L may be a carbon-carbon bond, but is preferably a functional group-bonded from the viewpoint of simplicity of synthesis.
- the bond by the functional group various types such as an ether bond, an ester bond, an amide bond, and a bond by phosphoric acid can be selected, and the amide bond is preferable.
- Preferred compounds of the general formula (I) for the GG mismatch in the Mismatchi base recognition molecule of the present invention include the following general formulas (II):
- R and R are a hydrogen atom, an alkyl group having 1 to 15 carbon atoms, and one or more carbon atoms in the alkyl group may be substituted with an oxygen atom or a nitrogen atom.
- R 2 is an alkyl group having 1 to 20 carbon atoms, and one or more carbon atoms in the alkyl group may be substituted with an oxygen atom, a nitrogen atom, or a carbonyl group. Represents a kill group. )
- immobilized product refers to a compound in which the above-mentioned compound is immobilized on a carrier or a compound in which the above-mentioned “branch” is extended so that it can be immobilized.
- the alkyl group for R 2 is a divalent alkyl group as shown in the general formula (II).
- mismatch base recognition molecule of the present invention can be used alone, it can be extended at an appropriate position in the molecule, for example, for immobilization from a linker part or linker.
- Labels can be used by introducing a radioactive element into a branch or the like, or by introducing a molecular species that emits chemiluminescence or fluorescence. Labeling as a measuring means can also be performed by labeling a nucleic acid portion such as DNA or RNA to be detected.
- it can be used by immobilizing it by bonding directly or using an alkylene group or the like to a polymer material such as polystyrene at an appropriate position of the Misuchichi base recognition molecule of the present invention.
- the Mismatsu base recognizing molecule of the present invention is a low molecular weight organic compound and can be appropriately produced by a usual organic synthesis method.
- the above-mentioned 1,8-naphthyridine derivative is capable of reacting 2-amino-1,8-naphthyridine or 2-amino-7-methyl-1,8-naphthyridine with N-protected 4-amino-butyric acid.
- a derivative such as an acid chloride can be reacted to acylate the amino group at the 2-position, and then the amino group can be produced by removing the protecting group.
- an amino protecting group used in peptide synthesis such as a hydrochloride, an acyl group or an alkoxycarbonyl group can be used.
- a target mismatch base recognition molecule By reacting the base recognition site thus obtained with a linker compound having a carboxyl group or a reactive derivative group at both ends, a target mismatch base recognition molecule can be obtained. it can. At this time, when a reactive group such as a nitrogen atom is present in the molecule of the linker compound, the compound can be appropriately protected with the above-mentioned protecting group and used.
- the Mismatchi base recognition molecule of the present invention detects the Mismatch base pair.
- a reagent for detecting mismatched base pairs by combining with an appropriate carrier can also be used as a base pair generating agent for forming a pseudo base pair from a base pair that cannot form a normal base pair.
- “pseudo base pair” means a base pair different from a naturally occurring base pair, and does not mean base pair strength.
- “normal base pair” refers to a naturally occurring base pair and refers to G—C, A—T, or A—U base pair.
- the present invention further relates to forming a normal base pair comprising the above-described mismatched base-recognizing molecule of the present invention and a material for detection and identification, for example, a material for chemiluminescence or fluorescence or a buffer solution.
- a material for detection and identification for example, a material for chemiluminescence or fluorescence or a buffer solution.
- a base pair that cannot form a normal base pair consisting of measuring the formation of the pseudo base pair is detected and identified. It provides a kit for
- the present invention uses the mismatched base recognition molecule of the present invention or the labeled or immobilized mismatched base recognition molecule of the present invention to detect, identify or quantify a pair of mismatched bases in DNA.
- a specific mismatched base pair for example, GG mismatch
- the base recognition site of the mismatch recognition molecule of the present invention forms a hydrogen bond with each of the bases in the specific mismatched base pair, and is stacked on a base pair existing in the vicinity of the base. This provides a DNA containing a mismatched base pair in which the mismatched base pair is stabilized.
- the mismatched base pair forms a “pair” (pseudo base pair) similar to the base pair by hydrogen bonding with the base recognition site of the mismatch recognition molecule of the present invention
- the mismatch recognition component of the present invention forming a “pair” with the mismatched base It is characterized in that the base recognition sites of the nucleotides are stacked sandwiched between bases forming adjacent, preferably adjacent base pairs in a sandwich manner.
- the mismatch recognition molecule of the present invention By using the mismatch recognition molecule of the present invention, it is possible to detect, identify, or quantify a mismatched base pair with high sensitivity and which cannot be achieved by conventional techniques. Since it forms stable and specific DNA, it can be applied to the treatment, prevention or diagnosis of various diseases associated with DNA damage. Further, since the DNA of the present invention can exist relatively stably in a state having mismatched base pairs, stabilization of DNA containing mismatched base pairs, cause of mismatch generation, mismatch It can also be used as research material for elucidation of the repair mechanism of spores.
- the present invention provides a method for hybridizing a single-stranded DNA or RNA serving as a specimen with a DNA or RNA having a normal nucleotide sequence corresponding thereto, and then normal base pairs in the hybridized DNA or RNA.
- a base pair that cannot form a base pair is formed into a pseudo-base pair with the base pair that cannot form a normal base pair, and
- a method for detecting a base pair that cannot form a normal base pair comprising measuring the formation of a target base pair, and a method for detecting a base sequence abnormality in DNA or RNA, comprising detecting and identifying a base pair. It also does.
- This method can be used when it is desired to check for the presence or absence of a gene abnormality.
- RNA a potentially abnormal DNA or its transcription product, RNA
- RNA is collected and hybridized with a complementary DNA or RNA having a normal nucleotide sequence to produce a double-stranded nucleic acid. If there is an abnormality in the base sequence of the collected gene, a mismatch will occur in the base pair at the base at the position of the abnormality.
- the mismatch recognition molecule of the present invention to the double-stranded nucleic acid in which the mismatch has occurred, the pseudo base pair described above is formed, and the presence or absence of the molecule in which the new pair is formed is measured. This allows simple and highly sensitive detection and identification of abnormalities in the collected genes.
- the mismatch recognition molecule of the present invention is a low-molecular organic compound, and when a new pair is formed, this molecule is incorporated into the nucleic acid. Therefore, both the unreacted mismatch recognition molecule of the present invention and the nucleic acids are used. It can be separated relatively easily.
- the mismatch recognition molecule of the present invention can be immobilized on a carrier and used.
- a mismatch recognition molecule specific to various mismatches of the present invention is immobilized on a plate such as a titer plate, and the above-described double-stranded nucleic acid.
- the labeled nucleic acid is added thereto, and the mixture is incubated for several minutes. After removal of the nucleic acids, the nucleic acids that specifically reacted with the mismatch recognition molecule of the present invention are trapped by the immobilized mismatch recognition molecule of the present invention, and can be detected and identified by labeling. Will be.
- the mismatch recognition molecule of the present invention can be immobilized on a metal thin film of a chip for detecting surface plasmon resonance (SPR).
- SPR surface plasmon resonance
- the presence or absence of a mismatch can be specifically detected simply by flowing the sample solution containing the double-stranded nucleic acid on the surface of the detection chip.
- mismatch recognition molecule of the present invention can be applied to many other detection means, and the present invention is not limited to these specific detection means.
- Example 1 Various concentrations of the compound obtained in Example 1 were added to the double-stranded DNA, and the DNA was examined by DNase I footprinting titration. That is, the DNA (Ku 4 nM scan Trang de concentration) of the double-stranded, N a C l (1 0 0 m M) and M g C l preparative squirrel-HCl buffer containing 2 (5 mM) (1 0 The mixture was incubated at 4 ° C for 12 hours together with the compound obtained in Example 1 adjusted to various concentrations with mM, pH 7.6). To this, 0.2 U of DN asel (DNA hydrolase) was added, and the mixture was incubated at 25 for 8 minutes. Thereafter, DNA was recovered by ethanol precipitation, and this was electrophoresed on a gel containing 12% polyacrylamide and 7 M urea.
- DN asel DNA hydrolase
- mismatch recognition molecule of the present invention By using the mismatch recognition molecule of the present invention, a mismatched base pair such as guanine-guanine mismatch, which cannot be achieved by conventional techniques, can be easily detected with high sensitivity.
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Description
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Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US09/889,976 US6607890B1 (en) | 1999-11-26 | 2000-10-26 | Mismatch-recognizing molecules |
DE60028556T DE60028556T2 (de) | 1999-11-26 | 2000-10-26 | Fehlpaarungen erkennende moleküle |
EP00970093A EP1164202B1 (en) | 1999-11-26 | 2000-10-26 | Mismatch-recognizing molecules |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP33662099A JP3705975B2 (ja) | 1999-11-26 | 1999-11-26 | ミスマッチ認識分子 |
JP11/336620 | 1999-11-26 |
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WO2001038571A1 true WO2001038571A1 (fr) | 2001-05-31 |
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PCT/JP2000/007497 WO2001038571A1 (fr) | 1999-11-26 | 2000-10-26 | Molecule reconnaissant les mesappariements |
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US (1) | US6607890B1 (ja) |
EP (1) | EP1164202B1 (ja) |
JP (1) | JP3705975B2 (ja) |
DE (1) | DE60028556T2 (ja) |
WO (1) | WO2001038571A1 (ja) |
Families Citing this family (11)
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JP3725405B2 (ja) | 2000-07-17 | 2005-12-14 | 独立行政法人科学技術振興機構 | テロメアなどに結合し得る分子、それを用いた方法 |
JP4011937B2 (ja) * | 2002-03-07 | 2007-11-21 | 独立行政法人科学技術振興機構 | ミスマッチ検出分子 |
JP4177006B2 (ja) * | 2002-03-08 | 2008-11-05 | 独立行政法人科学技術振興機構 | グアニンバルジ修飾分子 |
US20070037841A1 (en) * | 2003-02-28 | 2007-02-15 | Kazuhiko Nakatani | Inhibitors |
JP2004261083A (ja) * | 2003-02-28 | 2004-09-24 | Japan Science & Technology Agency | ミスマッチ認識分子およびミスマッチ検出方法、並びにその利用 |
JP4358579B2 (ja) * | 2003-02-28 | 2009-11-04 | 独立行政法人科学技術振興機構 | ミスマッチ検出分子およびミスマッチ検出方法、並びにその利用 |
JP2004325074A (ja) * | 2003-04-21 | 2004-11-18 | Japan Science & Technology Agency | ミスマッチ検出分子及びそれを用いたミスマッチの検出方法 |
JP4219748B2 (ja) * | 2003-06-24 | 2009-02-04 | 独立行政法人科学技術振興機構 | プローブの蛍光標識化方法 |
DE602005022990D1 (de) * | 2004-03-19 | 2010-09-30 | Japan Science & Tech Agency | Verfahren zum nachweis von genmutation und kit zum nachweis von genmutation |
JP4701378B2 (ja) * | 2004-10-07 | 2011-06-15 | 国立大学法人京都大学 | ミスマッチ塩基対検出分子およびミスマッチ塩基対検出方法、並びにその利用 |
JP5300335B2 (ja) * | 2008-06-06 | 2013-09-25 | 株式会社日清製粉グループ本社 | ミスマッチ塩基対含有核酸断片の高効率調製方法 |
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WO2000050424A1 (en) * | 1999-02-22 | 2000-08-31 | Biochem Pharma Inc. | [1,8] naphthyridine derivatives having antiviral activity |
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US6051942A (en) | 1996-04-12 | 2000-04-18 | Emerson Electric Motor Co. | Method and apparatus for controlling a switched reluctance machine |
JP4413325B2 (ja) | 1999-09-16 | 2010-02-10 | 独立行政法人科学技術振興機構 | バルジ塩基認識分子及びそれを含有するdna |
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1999
- 1999-11-26 JP JP33662099A patent/JP3705975B2/ja not_active Expired - Fee Related
-
2000
- 2000-10-26 DE DE60028556T patent/DE60028556T2/de not_active Expired - Lifetime
- 2000-10-26 US US09/889,976 patent/US6607890B1/en not_active Expired - Fee Related
- 2000-10-26 WO PCT/JP2000/007497 patent/WO2001038571A1/ja active IP Right Grant
- 2000-10-26 EP EP00970093A patent/EP1164202B1/en not_active Expired - Lifetime
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JP2000050876A (ja) * | 1998-08-11 | 2000-02-22 | Kyushu Univ | 遺伝子の特殊一本鎖核酸部位の検出用プローブ、遺伝子の特殊一本鎖核酸部位の検出方法およびその装置 |
WO2000050424A1 (en) * | 1999-02-22 | 2000-08-31 | Biochem Pharma Inc. | [1,8] naphthyridine derivatives having antiviral activity |
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MURRAY T.J. ET AL.: "7-Amido-1,8-naphthyridines as hydrogen bonding units for the complexation of guanine derivatives: the role of 2-alkoxyl group in decreasing binding affinity", TETRAHEDRON LETTERS, vol. 36, no. 42, 1995, pages 7627 - 7630, XP002936326 * |
NAKATANI K. ET AL.: "Recognition of a single guanine bulge by 2-acylamino-1,8-naphthyridine", J. AM. CHEM. SOC., vol. 122, no. 10, March 2000 (2000-03-01), pages 2172 - 2177, XP002936325 * |
SANDO S. ET AL.: "Scanning of guanine-guanine mismatches in DNA by synthetic ligands using surface plasmin resonance", NAT. BIOTECHNOL., vol. 19, no. 1, January 2001 (2001-01-01), pages 51 - 55, XP001004477 * |
See also references of EP1164202A4 * |
Also Published As
Publication number | Publication date |
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DE60028556D1 (de) | 2006-07-20 |
EP1164202A1 (en) | 2001-12-19 |
JP3705975B2 (ja) | 2005-10-12 |
DE60028556T2 (de) | 2007-05-16 |
EP1164202A4 (en) | 2005-01-05 |
EP1164202B1 (en) | 2006-06-07 |
US6607890B1 (en) | 2003-08-19 |
JP2001149096A (ja) | 2001-06-05 |
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