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1. 发明专利

JPS61109797A Labeled nucleotide and labeled polynucleotide 失效
标题翻译：标记核苷酸和标记的多核苷酸
公开(公告)号：JPS61109797A
公开(公告)日：1986-05-28
申请号：JP23184784
申请日：1984-11-01
申请人： Yuki Gosei Yakuhin Kogyo Kk
发明人： SUGIMOTO NOBUTAKA , SATO TOYOKI
IPC分类号： G01N33/53 , C07H19/04 , C07H19/14 , C07H19/23 , C07H21/04 , C12N15/00 , C12N15/09 , C12Q1/68 , G01N33/533 , G01N33/569
摘要： NEW MATERIAL:A compound bearing a labeled purine base analog of formula I (X
1 , X
2 , X
3 are C, N; Y, Z are H, hydroxyl, amino group; F is fluorescent labeling compound) where 7-deazapurine is excluded.
USE: A clinical diagnostic reagent used in identification of labeled genes, a reagent in biotechnology for extracting genes.
PREPARATION: For example, 2'-deoxyaribonucleoside transferase and thymidine are allowed to act on 4-amino-3(2-aminoethyl)-pyrazolo[3,4-d]pyrimidine and further phosphorus oxychloride is allowed to react with the product in anhydrous trimethyl phosphate to give 5'-monophosphate of formula III. The product is converted into 5'-triphosphate, which is used to effect nick translation whereby a polynucleotide bearing the group of formula I is obtained.
COPYRIGHT: (C)1986,JPO&Japio
摘要翻译：新材料：具有式I（X 1，X 2，X 3）的标记嘌呤碱基类似物的化合物是C，N; Y，Z是H，羟基，氨基; F是荧光标记化合物），其中7-脱氮嘌呤被排除。用途：用于鉴定标记基因的临床诊断试剂，用于提取基因的生物技术中的试剂。制备：例如，使2'-脱氧核糖核苷转移酶和胸苷作用于4-氨基-3（2-氨基乙基） - 吡唑并[3,4-d]嘧啶，进一步使磷酰氯与无水产物反应磷酸三甲酯，得到式III的5'-单磷酸酯。将产物转化为5'-三磷酸，其用于实现切口平移，由此获得具有式I基团的多核苷酸。

2. 发明专利

JPS6157595A Method of labeling polydeoxyribonucleotide or polyribonucleotide with an enzyme 失效
标题翻译：标记多核苷酸或多核苷酸与酶的方法
公开(公告)号：JPS6157595A
公开(公告)日：1986-03-24
申请号：JP17891684
申请日：1984-08-28
申请人： Yuki Gosei Yakuhin Kogyo Kk
发明人： SUGIMOTO NOBUTAKA
IPC分类号： C12Q1/68 , C07H21/02 , C07H21/04
摘要： PURPOSE: A poly(deoxy)ribonuleotide having a thiophosphate bond is covalently bonded with an enzyme at the thiol group in the thiophosphate to give a labeled compound which is suitable for identification of target genes.
CONSTITUTION: Polynucleotide such as polydeoxyribonucleotide or polyribonucleotide is allowed to react with, e.g., thiophosphoric acid or thiophosphoryl chloride to convert at least one of phosphate bonds into thiophosphate bonds. The resultant nucleotide is allowed to react with such an enzyme as it can form a covalent bond with the thiol group in the thiophosphate bond (which has the part of a crosslinking agent whose one end can react with the thiol group in the tiophosphate, because bearing, e.g., maleimide or dithio group and the other end can react with amino and mercapto groups in the enzyme and the other part of the enzyme bearing amino and mercapto groups such as a compound of the formula) at room temperature to give the objective compound.
COPYRIGHT: (C)1986,JPO&Japio
摘要翻译：目的：具有硫代磷酸键的聚（脱氧）核糖核苷酸与硫代磷酸酯硫醇基上的酶共价键合，得到适合于鉴定靶基因的标记化合物。构成：使多核苷酸如多脱氧核糖核苷酸或多核糖核苷酸与例如硫代磷酸或硫代磷酰氯反应，将至少一个磷酸键转化成硫代磷酸键。使所得核苷酸与这样的酶反应，因为它可以与硫代磷酸酯键中的硫醇基形成共价键（其具有一端可以与磷酸根中的硫醇基反应的交联剂的一部分，因为轴承，例如马来酰亚胺或二硫基，另一端可与酶和带有氨基和巯基的酶的其它部分如下式化合物的氨基和巯基在室温下反应，得到目标化合物。

3. 发明专利

JPH08298999A DETECTION OF VIROID OF POTATO STREPTOMYCES SCABIES AND PROBE FOR ITS DETECTION 失效
公开(公告)号：JPH08298999A
公开(公告)日：1996-11-19
申请号：JP12881995
申请日：1995-05-01
申请人： YUKI GOSEI YAKUHIN KOGYO KK
发明人： NAKAHARA KENJI , HATAYA TATSUJI , HAYASHI YOSHINORI , SUGIMOTO NOBUTAKA , KIMURA IKUO , YOMO EISHIRO
IPC分类号： G01N33/53 , C07H21/04 , C12N15/09 , C12Q1/68 , C12Q1/70
摘要： PURPOSE: To detect the potato Streptomyces scabies viroid(PSTVd) with high sensitivity for preventing potatoes from being infected with the viroids by using a plurality of synthetic oligonucleotides which are complementary to the PSTVd RNA as probes. CONSTITUTION: Synthetic oligonucleotides having bases sequences represented by formulas I to V which are complementary to the RNA of PSTVd are used in a mixture as probes. Raw leaves of tomato or the like infected with PSTVd are ground to extract the nucleic acid soluble in 2M lithium chloride, a reverse transcriptase is allowed to act on to synthesize cDNA, which is amplified by PCR. The amplified cDNA is immobilized to the microplate and a mixture of biotin-labeled synthetic oligonucleotides of formulas I to V is hybridized to the cDNA. Then, avidinated alkali phosphatase is allowed to react with the hybridomas and a color-developing substrate is allowed to act on the reaction mixture to determine PSTVd colorimetrically using a spectrophotometer.

4. 发明专利

JPH0231686A PRODUCTION OF TRIFLUOROTHYMIDINE 失效
公开(公告)号：JPH0231686A
公开(公告)日：1990-02-01
申请号：JP18174288
申请日：1988-07-22
申请人： YUKI GOSEI YAKUHIN KOGYO KK , JAPAN TOBACCO INC
发明人： SUGIMOTO NOBUTAKA , SUNAGA YONOSUKE , HAYASHI YOSHINORI , SHIBA TAKAMITSU , KATSURAYAMA AKIO , MIKAMI YOICHI
IPC分类号： C12P19/40 , C07H19/06 , C12P19/38 , C12R1/425
摘要： PURPOSE:To obtain trifluorothymidine in high yield by base exchange reaction under a mild temperature condition of 30-45 deg.C using bacteria classified as Serratia sp. between trifluoromethyl uracil and deoxyribose donor. CONSTITUTION:Bacterial classified as Serratia sp. such as Serratia marcescens IFO 3052 are put to culture in a conventional medium containing a carbon source such as glucose and a nitrogen source such as ammonium sulfate, and the cultured bacteria are separated from the resultant culture solution. Thence, a reaction is made at 30-45 deg.C in an aqueous solution containing the above bacteria and phosphoric acid between trifluoromethyl uracil and a deoxyribose donor selected from 2'-deoxyadenosine and 2'-deoxyinosine. This reaction may be made using an immobilized enzyme. After completing the reaction, the bacteria or enzyme is separated from the system, and the trifluorothymidine produced is isolated and purified. Thus, the objective trifluorothymidine can be obtained continuously in high yield.

5. 发明专利

JPH0223893A DETECTION OF HOP-STUNT VIROID AND DETECTION PROBE 失效
公开(公告)号：JPH0223893A
公开(公告)日：1990-01-26
申请号：JP2374688
申请日：1988-02-05
申请人： YUKI GOSEI YAKUHIN KOGYO KK
发明人： YOMO EISHIRO , SANO TERUO , KUDO HIROSHI , SUGIMOTO NOBUTAKA , SHIBA TAKAMITSU
IPC分类号： C12N15/09 , C12N15/00 , C12Q1/68
摘要： PURPOSE:To easily detect a viroid belonging to hop-stunt viroids by using a probe consisting of a synthetic oligonucleotide having complimentarity to the common sequence of viroid RNA. CONSTITUTION:A viroid belonging to the groups of hop-stunt viroid(HSV), hop-stunt viroid cucumber mutant, grape mutant or citrus mutant among viroids which are pathogens infecting to a plant and causing the symptoms such as growth inhibition or dwarf is hybridized by using a probe consisting of a synthetic oligo-nucleotide having complimintarity to the common sequence of the RNA of the above viroid, preferably a 15-25-mer synthetic oligonucleotide having the base sequence of formula. The hybridized product is detected by labeling with radioactive isotope, enzyme, fluorescence, etc.

6. 发明专利

JPS554308A ISOLATION OF DEOXYRIBONUCLEIC ACID FROM MILT OF FISH 失效
公开(公告)号：JPS554308A
公开(公告)日：1980-01-12
申请号：JP7540078
申请日：1978-06-23
申请人： YUKI GOSEI YAKUHIN KOGYO KK
发明人： SUGIMOTO NOBUTAKA , YOKOZAWA KUNIO , KISHIMOTO HIFUMI
IPC分类号： C07H1/08 , C07H21/04
摘要： PURPOSE:Homogenized milt of fish is made to a specific acidic value in pH to permit agitation without dilution with water and prevent the deterioration caused by deamination or others and followed by extraction with alkali, thus isolating DNA, an intermediate for drugs or food. CONSTITUTION:After homogenized, milt of fish is adjusted to 4.5-6.5 in pH by inorganic or organic acid and stirred at 80-100 deg.C for 0.5-2 hr. Then, an alkali aqueous solution is added to adjust the pH to over 8 and the stirring is continued, the solid part is separated and an acid is added to the filtrate to isolate deoxyribonucleic acid (DNA). The adjustment of the homogenized milt of fish to a specific pH value permits the increase in quantity to be treated per batch and the deterioration caused by deamination or others on heating can be prevented.

7. 发明专利

JPS6144352A Method for fluorescent marking of polydeoxyribonucleotide or polyribonucleotide 失效
标题翻译：氟代二氧化碳或聚氨酯的荧光标记方法
公开(公告)号：JPS6144352A
公开(公告)日：1986-03-04
申请号：JP16536184
申请日：1984-08-07
申请人： Yuki Gosei Yakuhin Kogyo Kk
发明人： KOBAYASHI MAKOTO , SUGIMOTO NOBUTAKA
IPC分类号： G01N33/50 , C07H21/00 , C07H21/04 , C12Q1/68 , G01N21/76 , G01N21/78 , G01N33/52 , G01N33/533
CPC分类号： C12Q1/68
摘要： PURPOSE:To enhance the detection sensitivity of marked polynucleotide by an optical marking method, by introducing an optical marking compound into the thiophosphoric acid group of a nucleic acid constitutional element. CONSTITUTION:The fluorescent marking of polynucleotide is performed by converting the phosphoric acid bond of polynucleotide to a thiophosphoric acid bond and reacting the thiol group of the thiophosphoric acid bond with a covalently bondable fluorescent marking compound specifically reacting said thiol group. The above-mentioned polynucleotide having the thiophosphoric acid bond can be synthesized by an org. synthesis method or enzymatic method. Especially, in the former org. synthesis method, it is easiest to introduce the thiophosphoric group into both terminal hydroxyl groups of polynucleotide and thiophosphoric acid, thiophosphoryl chloride or derivatives thereof may be reacted under a known condition.
摘要翻译：目的：通过光学标记法提高标记多核苷酸的检测灵敏度，通过将光学标记化合物引入核酸结构元件的硫代磷酸基团。构成：多核苷酸的荧光标记通过将多核苷酸的磷酸键转化为硫代磷酸键，并使硫代磷酸键的硫醇基与与所述硫醇基特别反应的共价键合的荧光标记化合物反应来进行。上述具有硫代磷酸键的多核苷酸可以通过组织合成。合成方法或酶法。特别是在前组织。合成方法中，最好将硫代磷酸引入多核苷酸和硫代磷酸的两个末端羟基，硫代磷酰氯或其衍生物可以在已知条件下反应。

8. 发明专利

JPS6157596A Method of labeling polydeoxyribonucleotide or polyribonucleotide with an enzyme 失效
标题翻译：标记多核苷酸或多核苷酸与酶的方法
公开(公告)号：JPS6157596A
公开(公告)日：1986-03-24
申请号：JP17891784
申请日：1984-08-28
申请人： Yuki Gosei Yakuhin Kogyo Kk
发明人： SUGIMOTO NOBUTAKA
IPC分类号： C12Q1/68 , C07H21/02 , C07H21/04
摘要： PURPOSE: DNA or RNA which has phosphonic acid bondage bearing a functional group as a part of phosphate bonds is allowed to react with a covalently bondable enzyme for labelling at the functional group whereby the titled compound labelled with an enzyme which is used for clinically diagnostic purposes with high detection sensitivity is obtained.
CONSTITUTION: The objective compound labelled with an enzyme is obtained by reaction of a polydeoxyribonucleotide or polyribonucleotide which has at least one of phosphoric bond bearing a functional group of the formula (R is 1W20C aliphatic hydrocarbon, aromatic hydrocarbon; X is a functional group which may be protected) among its phosphate bonds with a covalently bondable enzyme for labelling. The functional group linking to the phosphonic bond is, e.g., thiol or amino.
COPYRIGHT: (C)1986,JPO&Japio
摘要翻译：目的：将具有作为磷酸键的官能团的膦酸键的DNA或RNA与可共价键合的酶进行反应，以在官能团上进行标记，由此用用于临床诊断目的的酶标记的标题化合物获得高检测灵敏度。构成：用酶标记的目标化合物是通过具有至少一个具有下式官能团的磷键（R为1-20C脂肪族烃，芳族烃; X为官能团）的多脱氧核糖核苷酸或多核糖核苷酸其可以被保护）在其磷酸盐键之间，具有用于标记的共价键合酶。连接到膦酸键的官能团是例如硫醇或氨基。

9. 发明专利

JPS6144353A Method for fluorescent marking of polydeoxyribonucleotide or polyribonucleotide 失效
标题翻译：氟代二氧化碳或聚氨酯的荧光标记方法
公开(公告)号：JPS6144353A
公开(公告)日：1986-03-04
申请号：JP16536284
申请日：1984-08-07
申请人： Yuki Gosei Yakuhin Kogyo Kk
发明人： KOBAYASHI MAKOTO , SUGIMOTO NOBUTAKA
IPC分类号： G01N33/50 , C07H21/00 , C07H21/02 , C07H21/04 , C12Q1/00 , C12Q1/68 , G01N21/76 , G01N21/78 , G01N33/52 , G01N33/533 , G01N33/58
CPC分类号： C12Q1/68
摘要： PURPOSE:To enhance the detection sensitivity of marked polynucleotide by an optical marking method, by performing the fluorescent marking of phosphonic acid by reacting the functional group of a phosphonic acid group with an optical marking compound. CONSTITUTION:The fluorescent marking of polynucleotide is performed by converting the phosphoric acid group of polynucleotide to a phosphonic acid bond having a functional group represented by formula (wherein R is aliphatic or aromatic hydrocarbon which may have an 1-20C branched chain and X is a functional group which may have a protective group) and reacting said functional group with a covalently bondable fluorescent marking compound. As a reaction method of phosphonic acid and the fluorescent marking compound, there are a method wherein the fluorescent marking compound is directly bonded to a phosphorus atom through a phosphorus-carbon bond and a method wherein the functional group specifically reacted with the fluorescent marking compound is bonded to a phosphorus atom so as to interpose aliphatic or aromatic hydrocarbon, which may have a branched chain, therebetween and the fluorescent marking compound is subsequently reacted with said functional group but, from the aspect of reaction yield or reaction operation, the latter is more excellent.
摘要翻译：目的：通过光学标记方法提高标记多核苷酸的检测灵敏度，通过使膦酸基团的官能团与光学标记化合物反应来进行膦酸的荧光标记。构成：多核苷酸的荧光标记通过将多核苷酸的磷酸基团转换为具有由式（其中R为可具有1-20C支链的脂族或芳族烃，且X为可以具有保护基的官能团），并使所述官能团与共价键合的荧光标记化合物反应。作为膦酸和荧光标记化合物的反应方法，存在荧光标记化合物通过磷 - 碳键直接与磷原子键合的方法，其中与荧光标记化合物特异性反应的官能团为与磷原子键合以便在其间插入可以具有支链的脂族或芳族烃，并且荧光标记化合物随后与所述官能团反应，但是从反应产率或反应操作的方面来看，后者更多优秀。

10. 发明专利

JPS5740497A Preparation of 5-halogeno-2'-deoxyuridine 失效
标题翻译： 5-卤代二脱氧酶的制备
公开(公告)号：JPS5740497A
公开(公告)日：1982-03-06
申请号：JP11570580
申请日：1980-08-21
申请人： Yuki Gosei Yakuhin Kogyo Kk
发明人： SUGIMOTO NOBUTAKA , HOSOGAI MITSUO
IPC分类号： C07H19/073 , C07H19/06
摘要： PURPOSE: To obtain the titled compound useful as a medicine or an intermediate therefor, e.g. a remedy for cerebral tumor, in high yield, by reacting 2'-deoxyuridine with a halogenating agent in a nonaqueous organic solvent.
CONSTITUTION: 2'-Deoxyuridine of formula I is reacted with a halogenating agent in equimolar amounts in a nonaqueous organic solvent, e.g. pyridine, pyridine-acetic acid or DMF, at 20W70°C to give an intermediate product of formula II (X is halogen). The hydrogen halide is then eliminated from the compound of formula II continuously in the same solvent to afford the compound of formula III. 5-Bromo-2'- deoxyuridine is particularly useful as a remedy for cerebral tumor, and 5-iodo-2'- deoxyuridine is useful as an antiviral agent.
COPYRIGHT: (C)1982,JPO&Japio
摘要翻译：目的：获得用作药物或其中间体的标题化合物，例如。通过在非水有机溶剂中使2'-脱氧尿苷与卤化剂反应，以高产率获得脑瘤的治疗药物。构成：将式I的2'-脱氧尿苷与卤化剂以等摩尔量在非水有机溶剂中反应，吡啶，吡啶 - 乙酸或DMF，在20-70℃下反应，得到式II的中间产物（X为卤素）。然后在相同的溶剂中，从式II化合物中除去卤化氢，得到式III化合物。 5-溴-2'-脱氧尿苷特别可用作脑肿瘤的治疗剂，5-碘-2'-脱氧尿苷可用作抗病毒剂。

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