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1. 发明授权

US07399610B2 Method for cell-free protein synthesis using extract solution derived from insect cell 有权
标题翻译：使用衍生自昆虫细胞的提取物溶液进行无细胞蛋白质合成的方法
公开(公告)号：US07399610B2
公开(公告)日：2008-07-15
申请号：US11247294
申请日：2005-10-12
申请人： Masamitsu Shikata , Nobuhiro Hanafusa , Shinichiro Kobayashi
发明人： Masamitsu Shikata , Nobuhiro Hanafusa , Shinichiro Kobayashi
IPC分类号： C12P21/06
CPC分类号： C12P21/00
摘要： The present invention provides a simple cell-free protein synthesis method capable of affording synthesis of a protein in a high amount in a short time at a low cost. A method for cell-free protein synthesis using an extract derived from an insect cell, the method comprising removing a component which can pass through a semipermeable membrane through the semipermeable membrane while maintaining synthesis reaction, thereby to continuously synthesize a protein. Preferably, an mRNA is additionally supplied while said synthesis reaction is maintained. Further, said insect cell is preferably an established culture cell derived from Trichoplusia ni ovum cell.
摘要翻译：本发明提供了一种简单的无细胞蛋白质合成方法，其能够以低成本在短时间内高效地提供蛋白质的合成。一种使用来自昆虫细胞的提取物进行无细胞蛋白质合成的方法，该方法包括除去可以通过半透膜的半透膜的成分，同时保持合成反应，从而连续合成蛋白质。优选地，在保持所述合成反应的同时提供mRNA。此外，所述昆虫细胞优选是源自粉纹夜蛾卵母细胞的建立的培养细胞。

2. 发明申请

US20060121560A1 DNA fragment to promote translation reaction and method for cell-free protein synthesis system using the same 审中-公开
标题翻译： DNA片段促进翻译反应和方法，使用无细胞蛋白质合成系统
公开(公告)号：US20060121560A1
公开(公告)日：2006-06-08
申请号：US11295739
申请日：2005-12-07
申请人： Takashi Suzuki , Masaaki Ito , Toru Ezure , Masamitsu Shikata , Shinichiro Kobayashi
发明人： Takashi Suzuki , Masaaki Ito , Toru Ezure , Masamitsu Shikata , Shinichiro Kobayashi
IPC分类号： C12P21/06 , C12N5/06 , C12N15/63
CPC分类号： C12P21/00 , C07K14/43586
摘要： The present invention provides a DNA fragment allowing easy cloning of a desired gene and capable of further improving translation efficiency, a protein expression vector and a template DNA having the DNA fragment, a mRNA obtained from the template DNA, a reaction solution for cell-free protein synthesis system containing the template DNA or the mRNA, a method for cell-free protein synthesis system using the template DNA, and, kit for cell-free protein synthesis system including the expression vector. A DNA fragment having the base sequence represented by any of SEQ ID No. 1 to 11 to use for promoting translation reaction, a protein expression vector and a template DNA having the DNA fragment, a mRNA obtained from the template DNA, a reaction solution for cell-free protein synthesis system containing the template DNA or the mRNA, a method for cell-free protein synthesis system using the template DNA, and, kit for cell-free protein synthesis system including the expression vector.
摘要翻译：本发明提供了能够容易地克隆所需基因并能够进一步提高翻译效率的DNA片段，蛋白质表达载体和具有DNA片段的模板DNA，从模板DNA获得的mRNA，无细胞的反应溶液含有模板DNA或mRNA的蛋白质合成系统，使用模板DNA的无细胞蛋白质合成系统的方法，以及包含表达载体的无细胞蛋白质合成系统的试剂盒。具有用于促进翻译反应的SEQ ID No.1至11所示的碱基序列的DNA片段，具有该DNA片段的蛋白质表达载体和模板DNA，从模板DNA获得的mRNA，含有模板DNA或mRNA的无细胞蛋白质合成系统，使用模板DNA的无细胞蛋白质合成系统的方法，以及包含表达载体的无细胞蛋白质合成系统的试剂盒。

3. 发明申请

US20060084146A1 Method for cell-free protein synthesis using extract solution derived from insect cell 有权
标题翻译：使用衍生自昆虫细胞的提取物溶液进行无细胞蛋白质合成的方法
公开(公告)号：US20060084146A1
公开(公告)日：2006-04-20
申请号：US11247294
申请日：2005-10-12
申请人： Masamitsu Shikata , Nobuhiro Hanafusa , Shinichiro Kobayashi
发明人： Masamitsu Shikata , Nobuhiro Hanafusa , Shinichiro Kobayashi
IPC分类号： C12P21/06 , C12N5/06
CPC分类号： C12P21/00
摘要： The present invention provides a simple cell-free protein synthesis method capable of affording synthesis of a protein in a high amount in a short time at a low cost. A method for cell-free protein synthesis using an extract derived from an insect cell, the method comprising removing a component which can pass through a semipermeable membrane through the semipermeable membrane while maintaining synthesis reaction, thereby to continuously synthesize a protein. Preferably, an mRNA is additionally supplied while said synthesis reaction is maintained. Further, said insect cell is preferably an established culture cell derived from Trichoplusia ni ovum cell.
摘要翻译：本发明提供了一种简单的无细胞蛋白质合成方法，其能够以低成本在短时间内高效地提供蛋白质的合成。一种使用来自昆虫细胞的提取物进行无细胞蛋白质合成的方法，该方法包括除去可以通过半透膜的半透膜的成分，同时保持合成反应，从而连续合成蛋白质。优选地，在保持所述合成反应的同时提供mRNA。此外，所述昆虫细胞优选是源自粉纹夜蛾卵母细胞的建立的培养细胞。

4. 发明申请

US20070212752A1 Cell-free protein synthesis for controlling introduction of modification group into protein 审中-公开
标题翻译：用于控制将修饰基团导入蛋白质的无细胞蛋白质合成
公开(公告)号：US20070212752A1
公开(公告)日：2007-09-13
申请号：US11700823
申请日：2007-02-01
申请人： Takashi Suzuki , Masaaki Ito , Toru Ezure , Masamitsu Shikata , Shinichiro Kobayashi , Toshihiko Utsumi
发明人： Takashi Suzuki , Masaaki Ito , Toru Ezure , Masamitsu Shikata , Shinichiro Kobayashi , Toshihiko Utsumi
IPC分类号： C12P21/06 , C12N5/06 , C07K14/47
CPC分类号： C12P21/02
摘要： It is intended to provide a method capable of introducing an arbitrary modification group into a synthetic protein and a method capable of easily controlling the syntheses of a modified protein and an unmodified protein. The present invention provides a method of cell-free protein synthesis comprising: performing cell-free protein synthesis using as a reaction solution of cell-free protein synthesis, a mixture solution of an extract derived from a eukaryotic cell and a reagent solution comprising as a substrate, a substance having a desired group to be introduced as a post-translational modification group; and thereby obtaining a protein comprising the post-translational modification group introduced therein. Preferably, the group to be introduced is an acyl group of fatty acid, and the form of the post-translational modification is lipid modification at the N-terminus of the protein.
摘要翻译：旨在提供能够将任意修饰基团引入合成蛋白质的方法和能够容易地控制修饰蛋白质和未修饰蛋白质的合成的方法。本发明提供了一种无细胞蛋白质合成方法，包括：使用无细胞蛋白质合成的反应溶液进行无细胞蛋白质合成，来自真核细胞的提取物的混合溶液和包含作为底物，具有作为翻译后修饰基团引入的所需基团的物质; 从而获得包含其中引入的翻译后修饰基团的蛋白质。优选地，待引入的基团是脂肪酸的酰基，并且翻译后修饰的形式是蛋白质N末端的脂质修饰。

5. 发明授权

US08697400B2 Method for amplification of DNA from blood sample and DNA amplification kit therefor 有权
标题翻译：从血液样品中扩增DNA的方法及其DNA扩增试剂盒
公开(公告)号：US08697400B2
公开(公告)日：2014-04-15
申请号：US12637239
申请日：2009-12-14
申请人： Masamitsu Shikata
发明人： Masamitsu Shikata
IPC分类号： C12P19/34
CPC分类号： C12Q1/6846 , C12Q2537/119 , C12Q2527/119
摘要： A DNA amplification method including: subjecting a blood sample having DNA to be amplified, to a pretreatment using an alkaline aqueous solution under ordinary temperature, so as to extract double-stranded DNA from the blood sample and dissociate the double-stranded DNA into a single-stranded DNA to obtain a blood-derived sample including the single-stranded DNA; preparing an isothermal amplification reaction solution comprising a mixture of the blood-derived sample, a primer, dNTP, a strand-displacing DNA polymerase, a magnesium salt and a buffer, to establish an isothermal amplification reaction system meeting optimum conditions for the strand-displacing DNA polymerase; and amplifying DNA in the isothermal amplification reaction system using the single-stranded DNA as a template.
摘要翻译：一种DNA扩增方法，其特征在于，将通过DNA扩增的血液样品在常温下使用碱性水溶液进行预处理，从血液样品中提取双链DNA，将双链DNA解离为单一DNA 品种DNA以获得包含单链DNA的血液衍生物样品; 制备包含血液衍生的样品，引物，dNTP，链置换DNA聚合酶，镁盐和缓冲液的混合物的等温扩增反应溶液，以建立满足链取代最佳条件的等温扩增反应系统 DNA聚合酶; 并使用单链DNA作为模板在等温扩增反应体系中扩增DNA。

6. 发明申请

US20100151531A1 METHOD FOR AMPLIFICATION OF DNA FROM BLOOD SAMPLE AND DNA AMPLIFICATION KIT THEREFOR 有权
标题翻译：用于从血液样品和DNA扩增试剂盒中放大DNA的方法
公开(公告)号：US20100151531A1
公开(公告)日：2010-06-17
申请号：US12637239
申请日：2009-12-14
申请人： Masamitsu SHIKATA
发明人： Masamitsu SHIKATA
IPC分类号： C12P19/34 , C12N9/96
CPC分类号： C12Q1/6846 , C12Q2537/119 , C12Q2527/119
摘要： A DNA amplification method including: subjecting a blood sample having DNA to be amplified, to a pretreatment using an alkaline aqueous solution under ordinary temperature, so as to extract double-stranded DNA from the blood sample and dissociate the double-stranded DNA into a single-stranded DNA to obtain a blood-derived sample including the single-stranded DNA; preparing an isothermal amplification reaction solution comprising a mixture of the blood-derived sample, a primer, dNTP, a strand-displacing DNA polymerase, a magnesium salt and a buffer, to establish an isothermal amplification reaction system meeting optimum conditions for the strand-displacing DNA polymerase; and amplifying DNA in the isothermal amplification reaction system using the single-stranded DNA as a template.
摘要翻译：一种DNA扩增方法，其特征在于，将通过DNA扩增的血液样品在常温下使用碱性水溶液进行预处理，从血液样品中提取双链DNA，将双链DNA解离为单一DNA 品种DNA以获得包含单链DNA的血液衍生物样品; 制备包含血液衍生的样品，引物，dNTP，链置换DNA聚合酶，镁盐和缓冲液的混合物的等温扩增反应溶液，以建立满足链取代的最佳条件的等温扩增反应系统 DNA聚合酶; 并使用单链DNA作为模板在等温扩增反应体系中扩增DNA。

7. 发明申请

US20090155881A1 CELL-FREE PROTEIN SYNTHESIS METHOD AND CELL-FREE PROTEIN SYNTHESIS REACTION SOLUTION USING ADENOSINE 3',5'-BISPHOSPHATE 审中-公开
标题翻译：无细胞蛋白合成方法和无细胞蛋白合成反应溶液使用腺苷3'，5'-二磷酸
公开(公告)号：US20090155881A1
公开(公告)日：2009-06-18
申请号：US12323233
申请日：2008-11-25
申请人： Masamitsu Shikata , Masaaki Ito , Toru Ezure , Takashi Suzuki
发明人： Masamitsu Shikata , Masaaki Ito , Toru Ezure , Takashi Suzuki
IPC分类号： C12N9/12
CPC分类号： C12P21/02
摘要： The present invention provides a method of conducting cell-free protein synthesis by conveniently suppressing mRNA degradation, and a reaction solution enabling cell-free protein synthesis by conveniently suppressing mRNA degradation. A cell-free protein synthesis method using a cell-free protein synthesis reaction solution containing at least an extract liquid derived from a living cell, a potassium salt, a magnesium salt, adenosine triphosphate, guanosine triphosphate, creatine phosphate, creatine kinase, amino acid, a tRNA, an mRNA, a buffer, and adenosine 3′,5′-bisphosphate.
摘要翻译：本发明提供通过方便地抑制mRNA降解进行无细胞蛋白质合成的方法，以及通过方便地抑制mRNA降解使得能够进行无细胞蛋白质合成的反应溶液。一种无细胞蛋白质合成方法，其使用至少含有来自活细胞，钾盐，镁盐，三磷酸腺苷，三磷酸鸟苷，肌酸磷酸肌酸，肌酸激酶，氨基酸的提取液的无细胞蛋白质合成反应溶液，tRNA，mRNA，缓冲液和腺苷3'，5'-二磷酸盐。

8. 发明申请

US20070128688A1 Method for cell-free protein synthesis using complementary oligonucleotide 审中-公开
标题翻译：使用互补寡核苷酸的无细胞蛋白质合成方法
公开(公告)号：US20070128688A1
公开(公告)日：2007-06-07
申请号：US11604334
申请日：2006-11-27
申请人： Masamitsu Shikata , Shinichiro Kobayashi
发明人： Masamitsu Shikata , Shinichiro Kobayashi
IPC分类号： C12P21/06 , C12N5/06
CPC分类号： C12P21/00
摘要： The present invention provides a practical cell-free protein synthesis method capable of easily synthesizing a large amount of protein at low cost. A method for cell-free protein synthesis performed in a reaction solution containing mRNA and a living cell-derived extract solution, the reaction solution containing an oligonucleotide complementary to a sequence present in a 3′ terminal region of the mRNA.
摘要翻译：本发明提供了一种能够以低成本容易地合成大量蛋白质的实用的无细胞蛋白质合成方法。在含有mRNA和活细胞衍生的提取液的反应液中进行无细胞蛋白质合成的方法，所述反应液含有与存在于mRNA的3'端区域的序列互补的寡核苷酸。

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