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1. 发明专利

JP2000197493A PREPARATION OF DOUBLE-STRANDED DNA EXCEPT A PART OF TERMINAL HOMOPOLYMER AND DETERMINATION OF BASE SEQUENCE THEREOF 失效
公开(公告)号：JP2000197493A
公开(公告)日：2000-07-18
申请号：JP31503699
申请日：1999-11-05
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12N15/09 , C12Q1/68
摘要： PROBLEM TO BE SOLVED: To obtain a double-stranded DNA that is useful for decision the base sequence by allowing a restriction enzyme having the cleavage site outside the recognition site to act on the double-stranded DNA having a homopolymer on the chain end and removing the homopolymer part. SOLUTION: A restriction enzyme having the cleavage site on the outside of the recognition site is allowed to act on a double stranded DNA bearing homopolymer parts on one or both chain ends and at least one of the homopolymer is cut off to obtain the objective double stranded DNA from which the homopolymers that disturbs the decision of the base sequence and locating on the chain ends are selectively removed wholly or partially and is useful as a template for deciding the base sequence. The homopolymer part comprises the homopolymer and non-homopolymer and at least one part of the non-homopolymer is the recognition site of the restriction enzyme and the restriction enzyme is preferably in the class IIS.

2. 发明专利

JP2000146958A METHOD FOR IMMOBILIZING AND SUPPORTING DNA AND SUPPORT FOR IMMOBILIZING DNA 失效
公开(公告)号：JP2000146958A
公开(公告)日：2000-05-26
申请号：JP24091099
申请日：1999-08-27
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12N15/09 , C12M1/00 , C12Q1/68 , G01N33/48 , G01N33/50 , G01N33/53 , G01N33/566 , G01N37/00
摘要： PROBLEM TO BE SOLVED: To enable the speedy and efficient storage and distribution of DNA without the need for time and effort. SOLUTION: By adhering a DNA solution to a sheet-shaped support 10 with a predetermined thickness and drying the DNA solution adhered to the support 10, DNA 12 is immobilized to the support 10. As it is possible to immobilize or print DNA to the sheet-shaped support 10 by the simple operation of adhering the DNA solution to the sheet-shaped support 10 and drying it, it becomes possible to store and distribute DNA speedily and efficiently without the need for time effort.

3. 发明专利

JP2000081417A CONDUCTIVE DISSOLVING AND REMOVING METHOD FOR GEL PRESENT IN TUBULAR CONTAINER 失效
公开(公告)号：JP2000081417A
公开(公告)日：2000-03-21
申请号：JP25059898
申请日：1998-09-04
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12M1/00 , G01N27/447
摘要： PROBLEM TO BE SOLVED: To remove a gel remaining in a tubular container such as a capillary or mold without damaging or dismantling the tubular container. SOLUTION: This conductive dissolving and removing method conductively dissolves and removes a gel generated by polymerizing at least acrylamide or a derivative of acrylamide or a derivative mixture of acrylamide, a crosslinking agent, and a polymerizing agent together in a tubular container. In this case, using an electrolyte containing a component solating the gel, electricitiy is continuously applied to each end of the gel present in the tubular container, whereby the dynamic strength of the get present in the tubular container is decreased.

4. 发明专利

JP2000035432A ORIGONUCLEOTIDE DETECTING METHOD AND BASE SEQUENCE DETERMINATION METHOD 失效
公开(公告)号：JP2000035432A
公开(公告)日：2000-02-02
申请号：JP20361998
申请日：1998-07-17
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： G01N33/58 , B01L3/00 , C12N15/09 , C12Q1/68 , G01N27/447
摘要： PROBLEM TO BE SOLVED: To provide a method and its device by which plural samples are simultaneously analyzed so that a base sequence having large quantities of genetic information can be determined in a short time. SOLUTION: Oligonucleotide segments derived from a sample nucleic acid of each of plural nucleic acid samples are segmented so that four kinds of oligonucleotide segments, which have four kinds of terminal bases respectively, are prepared. Each the sample nucleic acid is labelled with a specific label different from the other, and then, is analyzed by a method by which each the oligonucleotide segment can be identified according to difference of a length of one base. Thus, a base sequence of a target nucleic acid is decided.

5. 发明专利

JPH11137296A DETERMINATION OF BASE SEQUENCE OF DNA 失效
公开(公告)号：JPH11137296A
公开(公告)日：1999-05-25
申请号：JP30762797
申请日：1997-11-10
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12N15/09 , C12Q1/68
摘要： PROBLEM TO BE SOLVED: To provide a method for favorably determining the base sequence of a DNA without inhibiting its transcription due to RNA polymerase even in the case of using a compression inhibition ribonucleotide derivative as a ribonucleotide in order to improving sequence reading accuracy. SOLUTION: This method for determining the base sequence of a DNA comprises the following procedure: in the presence of at DNA fragment containing an RNA polymerase and a promoter sequence therefor, a ribonucleoside-5-triphosphate is reacted with 3-deoxyribonucleoside-5-triphosphate to form a nucleic acid-transcribed product, which is then separated, and from the resulting fraction, the aimed sequence of the nucleic acid is read out; wherein at least one kind of nucleic acid transcription initiator selected from oligoribonucleotides bearing no phosphate group on the 5-terminal and oligoribonucleotides bearing mono- or diphosphate group on the 5-terminal is used.

6. 发明专利

JP2001027633A METHOD FOR DETECTING INTERACTING PROTEIN 失效
公开(公告)号：JP2001027633A
公开(公告)日：2001-01-30
申请号：JP22426999
申请日：1999-08-06
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12N15/09 , C07K1/13 , C07K17/00 , C12P21/02 , C40B30/04 , G01N33/15 , G01N33/50 , G01N33/566 , G01N33/68
摘要： PROBLEM TO BE SOLVED: To easily and speedily detect an interaction of proteins by synthesizing and mixing a protein having a label for detection and a protein having a modification for separation by a cell-frame protein synthetic method, separating a protein couple formed by an interaction, and identifying the protein couple with the utilization of the label for detection. SOLUTION: A protein having a label for detection and a protein having a modification for separation are synthesized by a cell-free protein synthesis method. A fluorescent substance, chemical emission substance, radioactive isotope or the like is used as the label for detection, and forming biotin, forming avidin, forming streptoavidin or the like is used as the modification for separation. After synthesized, the proteins are mixed to interact with each other. The formed protein couple has the label for detection and modification for separation. The protein having the label for detection which does not form the protein couple is separated with the utilization of a function of the modification for separation. At this time, the protein having the modification for separation which does not form the protein couple is also separated simultaneously, so that only the protein couple having the label for detection and modification for separation can be detected.

7. 发明专利

JPH11206400A DETERMINATION OF BASE SEQUENCE OF DNA 失效
公开(公告)号：JPH11206400A
公开(公告)日：1999-08-03
申请号：JP1047198
申请日：1998-01-22
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12N15/09 , C12Q1/68
摘要： PROBLEM TO BE SOLVED: To provide a method for determining the base sequence of a DNA utilizing a new method for simultaneously enabling the amplification of a target DNA and the production of a nucleic acid transcript without using a thermostable RNA polymerase. SOLUTION: This method is to determine the base sequence of a target DNA fragment and comprises producing a nucleic acid transcript in the presence of a terminator for the nucleic acid transription reaction with an RNA polymerase using an amplified target DNA fragment as a template while amplifying the target DNA fragment and analyzing the produced nucleic acid transcript. The amplification of the target DNA fragment and the production of the nucleic acid transcript are carried out under isothermal conditions. The amplification of the target DNA fragment and the production of the nucleic acid transcript are conducted at about normal temperatures and the amplification of the target DNA fragment can be carried out by a strand displacement amplification method.

8. 发明专利

JP2002360256A METHOD FOR PRODUCING RNA PROBE, METHOD FOR DETECTING TARGET NUCLEIC ACID, AND KIT FOR PRODUCING RNA PROBE 失效
公开(公告)号：JP2002360256A
公开(公告)日：2002-12-17
申请号：JP2001167910
申请日：2001-06-04
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE , OKAZAKI YASUSHI
IPC分类号： G01N33/53 , C12N15/09 , C12Q1/68 , G01N33/566 , G01N37/00
摘要： PROBLEM TO BE SOLVED: To provide a method for producing a labeled RNA probe including a fluorescent label and capable of providing a high S/N ratio, and further to provide a method for detecting a target nucleic acid, and a kit for producing the RNA probe. SOLUTION: This method for producing the RNA probe having the label comprises reacting an RNA polymerase in the presence of a DNA fragment including a promoter sequence, and a substrate for the RNA polymerase. At least one of the substrates has the label, and the RNA polymerase is a variant RNA polymerase obtained by modifying at least one amino acid of a wild-type RNA polymerase so as to be able to take the substrate having the label in. The method for detecting the target nucleic acid comprises selectively detecting the RNA probe hybridized with the target nucleic acid by mixing the target nucleic acid with the RNA probe produced by the method and having the label. The kit for producing the RNA probe is also provided.

9. 发明专利

JP2002253237A METHOD OF PREPARING NORMALIZED AND/OR SUBTRACTED COMPLEMENTARY DNA LIBRARY 审中-公开
公开(公告)号：JP2002253237A
公开(公告)日：2002-09-10
申请号：JP2001256576
申请日：2001-08-27
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12N15/09
摘要： PROBLEM TO BE SOLVED: To provide a method of efficiently preparing normalized and/or subtracted long-chain and full-length coding or full length cDNA libraries. SOLUTION: A method of preparing normalized and/or subtracted cDNA wherein the normalized and/or subtracted cDNA is the reverse transcript of mRNA, namely uncloned cDNA (cDNA tester), is provided. The method of preparing the cDNA libraries comprises the step (a) is which the cDNA tester is prepared; the step (b) in which the normalized and/or subtracted RNA driver are prepared; the step (c) where the resultant normalized and/or subtracted RNA driver are mixed with the cDNA tester; the step (d) in which an enzyme capable of cleaving the single strand sites among the normalized and/or subtracted drivers nonspecifically binding to the cDNA tester; the step (e) in which the single strand RNA driver cleaved is removed from the tester and the tester/driver hybrids are also removed; and the step (f) in which the objective normalized and/or subtracted cDNA are collected.

10. 发明专利

JP2001352974A METHOD FOR ACTIVATING ENZYME AT HIGH TEMPERATURE 失效
公开(公告)号：JP2001352974A
公开(公告)日：2001-12-25
申请号：JP2001128132
申请日：2001-04-25
申请人： RIKAGAKU KENKYUSHO , HAYASHIZAKI YOSHIHIDE
发明人： HAYASHIZAKI YOSHIHIDE
IPC分类号： C12N9/00 , C12N9/12
摘要： PROBLEM TO BE SOLVED: To provide a method for more easily and effectively improving the heat resistance of an enzyme and thereby manifesting high enzymatic activity at high temperatures. SOLUTION: This method for enhancing enzymatic activity at high temperatures comprises including, in a reaction liquid containing a relevant enzyme, substance(s) having chaperonic action, e.g. saccharide(s) such as trehalose, sorbitol and/or arabitol, amino acid(s) such as betaine and/or sarcosine, chapenonin-like protein.

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