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1. 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。

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

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。

4. 发明授权

US07745171B2 Cell-free protein synthesis method using insect cell extract solution 有权
标题翻译：无细胞蛋白质合成方法采用昆虫细胞提取液
公开(公告)号：US07745171B2
公开(公告)日：2010-06-29
申请号：US11987981
申请日：2007-12-06
申请人： Toru Ezure , Shoken Higashide , Takashi Suzuki , Masaaki Ito , Koki Endo
发明人： Toru Ezure , Shoken Higashide , Takashi Suzuki , Masaaki Ito , Koki Endo
IPC分类号： C12P21/06 , C07K1/00
CPC分类号： C12N1/066 , C12P21/00
摘要： The present invention provides a preparation method of an insect cell extract solution for cell-free protein synthesis, the insect cell extract solution, a protein synthesis method in a cell-free system, which uses the insect cell extract solution, and a kit for cell-free protein synthesis containing the insect cell extract solution. The extract solution is easily prepared by the method of the present invention and can synthesize a higher amount of protein than by extract solutions prepared by conventional methods.
摘要翻译：本发明提供了一种用于无细胞蛋白质合成的昆虫细胞提取液的制备方法，昆虫细胞提取液，使用昆虫细胞提取液的无细胞系统中的蛋白质合成方法，以及细胞用试剂盒 - 含有昆虫细胞提取液的蛋白质合成。提取液可以通过本发明的方法容易地制备，并且可以通过常规方法制备的提取物溶液合成更多的蛋白质。

5. 发明申请

US20060141559A1 Extract from cultured mammalian cell, process for preparation thereof and method of cell-free protein synthesis using the extract 审中-公开
标题翻译：培养的哺乳动物细胞的提取物，其制备方法和使用提取物的无细胞蛋白质合成方法
公开(公告)号：US20060141559A1
公开(公告)日：2006-06-29
申请号：US10559500
申请日：2004-06-09
申请人： Takashi Suzuki , Toru Ezure , Masaaki Ito , Shoken Higashide , Koki Endo
发明人： Takashi Suzuki , Toru Ezure , Masaaki Ito , Shoken Higashide , Koki Endo
IPC分类号： C12P21/06 , C12N5/06
CPC分类号： C12P21/02
摘要： A method for preparing a cultured mammalian cell extract liquid, comprising at least the step of rapidly freezing a cultured mammalian cell suspended in a solution for extraction; a cultured mammalian cell extract liquid prepared by the process; and a method for cell-free protein synthesis using the extract liquid. Preferably, the method for cell-free protein synthesis comprises the step of prior to conducting of synthetic reaction, effecting incubation of a reaction liquid for cell-free protein synthesis in a state of containing components other than exogenous mRNA for a specified period of time.
摘要翻译：一种制备培养的哺乳动物细胞提取液的方法，至少包括将悬浮于溶液中的培养的哺乳动物细胞迅速冷冻以提取的步骤; 通过该方法制备的培养的哺乳动物细胞提取液; 以及使用提取液的无细胞蛋白质合成方法。优选地，无细胞蛋白质合成的方法包括在进行合成反应之前，在含有外源mRNA以外的成分的状态下培养反应液进行无细胞蛋白质合成的步骤达规定时间。

6. 发明授权

US07387884B2 Yeast extract solution for cell-free protein synthesis, method for preparation thereof and method for cell-free protein synthesis using same 有权
标题翻译：用于无细胞蛋白质合成的酵母提取物溶液，其制备方法和使用其的无细胞蛋白质合成方法
公开(公告)号：US07387884B2
公开(公告)日：2008-06-17
申请号：US10751962
申请日：2004-01-07
申请人： Takashi Suzuki , Toru Ezure , Masaaki Ito
发明人： Takashi Suzuki , Toru Ezure , Masaaki Ito
IPC分类号： C12P21/06
CPC分类号： C12P21/02 , C12N1/063
摘要： The present invention provides a preparation method of a yeast extract solution for cell-free protein synthesis, which solution is easy to prepare and is capable of synthesizing a higher amount of a protein than by conventional yeast extract solutions, the yeast extract solution, a cell-free synthesis method of protein, which uses the yeast extract solution, and a kit for cell-free protein synthesis containing the yeast extract solution. The method of the present invention includes rupturing a yeast cell in a frozen state, and obtaining an extract thereof.
摘要翻译：本发明提供了一种用于无细胞蛋白质合成的酵母提取物溶液的制备方法，该溶液易于制备并且能够合成比常规酵母提取物溶液更高量的蛋白质，酵母提取物溶液，细胞 - 使用酵母提取物溶液的蛋白质的合成方法和含有酵母提取物溶液的无细胞蛋白质合成试剂盒。本发明的方法包括在冷冻状态下破裂酵母细胞，并获得其提取物。

7. 发明申请

US20080221311A1 Preparation method of insect cell extract solution for cell-free protein synthesis, the insect cell extract solution and cell-free synthesis method of protein using the insect cell extract solution 有权
公开(公告)号：US20080221311A1
公开(公告)日：2008-09-11
申请号：US11987981
申请日：2007-12-06
申请人： Toru Ezure , Shoken Higashide , Takashi Suzuki , Masaaki Ito , Koki Endo
发明人： Toru Ezure , Shoken Higashide , Takashi Suzuki , Masaaki Ito , Koki Endo
IPC分类号： C07K1/00
CPC分类号： C12N1/066 , C12P21/00
摘要： The present invention provides a preparation method of an insect cell extract solution for cell-free protein synthesis, the insect cell extract solution, a protein synthesis method in a cell-free system, which uses the insect cell extract solution, and a kit for cell-free protein synthesis containing the insect cell extract solution. The extract solution is easily prepared by the method of the present invention and can synthesize a higher amount of protein than by extract solutions prepared by conventional methods.

8. 发明授权

US07323332B2 Preparation method of insect cell extract solution for cell-free protein synthesis, the insect cell extract solution and cell-free synthesis method of protein using the insect cell extract solution 有权
标题翻译：用于无细胞蛋白质合成的昆虫细胞提取液的制备方法，昆虫细胞提取液和无细胞合成方法的蛋白质使用昆虫细胞提取液
公开(公告)号：US07323332B2
公开(公告)日：2008-01-29
申请号：US10742766
申请日：2003-12-23
申请人： Toru Ezure , Shoken Higashide , Takashi Suzuki , Masaaki Ito , Koki Endo
发明人： Toru Ezure , Shoken Higashide , Takashi Suzuki , Masaaki Ito , Koki Endo
IPC分类号： C12N5/06 , C12P21/06
CPC分类号： C12N1/066 , C12P21/00
摘要： The present invention provides a preparation method of an insect cell extract solution for cell-free protein synthesis, the insect cell extract solution, a protein synthesis method in a cell-free system, which uses the insect cell extract solution, and a kit for cell-free protein synthesis containing the insect cell extract solution. The extract solution is easily prepared by the method of the present invention and can synthesize a higher amount of protein than by extract solutions prepared by conventional methods.
摘要翻译：本发明提供了一种用于无细胞蛋白质合成的昆虫细胞提取液的制备方法，昆虫细胞提取液，使用昆虫细胞提取液的无细胞系统中的蛋白质合成方法，以及细胞用试剂盒 - 含有昆虫细胞提取液的蛋白质合成。提取液可以通过本发明的方法容易地制备，并且可以通过常规方法制备的提取物溶液合成更多的蛋白质。

9. 发明授权

US07189528B2 Extract solution for cell-free protein synthesis, method for cell-free protein synthesis using same and method for preparation of the extract solution 失效
标题翻译：无细胞蛋白质合成提取液，无细胞蛋白质合成方法及提取液的制备方法
公开(公告)号：US07189528B2
公开(公告)日：2007-03-13
申请号：US10315108
申请日：2002-12-10
申请人： Shoken Higashide , Toru Ezure , Satoko Shiraki , Masaaki Ito
发明人： Shoken Higashide , Toru Ezure , Satoko Shiraki , Masaaki Ito
IPC分类号： C12P21/06
CPC分类号： C12N9/0069 , C07K14/43586
摘要： The present invention provides a novel extract solution for cell-free protein synthesis, which contains at least an extract derived from a Bombyx mori L. tissue and an exogenous mRNA, which is easy to prepare and which is capable of realizing cell-free protein synthesis that can synthesize glycoprotein, a cell-free protein synthesis method using it, and a method for preparing an extract solution for cell-free protein synthesis, which includes adding an exogenous mRNA to an extract derived from a Bombyx mori L. tissue.
摘要翻译：本发明提供了一种新型的无细胞蛋白质合成提取液，其至少含有来源于蚕蛾组织的提取物和易于制备的能够实现无细胞蛋白质合成的外源mRNA 可以合成糖蛋白，使用其的无细胞蛋白质合成方法，以及制备无细胞蛋白质合成提取液的方法，其包括向来自家蚕组织的提取物中添加外源mRNA。

10. 发明申请

US20070099264A1 Method of posttranslational modification by adding mycrosomal membrane in cell-free protein synthesis 审中-公开
标题翻译：通过在无细胞蛋白质合成中添加霉菌膜进行翻译后修饰的方法
公开(公告)号：US20070099264A1
公开(公告)日：2007-05-03
申请号：US10578665
申请日：2004-11-12
申请人： Toshihiko Utsumi , Koki Endo , Masaaki Ito
发明人： Toshihiko Utsumi , Koki Endo , Masaaki Ito
IPC分类号： C12P21/06 , C07H21/04 , C12N5/06 , C07K14/47 , C07K14/78
CPC分类号： C12P21/005
摘要： It is intended to provide a novel method for posttranslational modification in cell-free protein synthesis and a novel method for cell-free protein synthesis with the use of such posttranslational modification reaction. A method for synthesizing a protein in a cell-free system using an extract liquid for cell-free protein synthesis containing an arthropod-derived extract, characterized in that translation reaction is carried out in the presence of mammal-derived microsomal membranes.
摘要翻译：旨在提供一种用于无细胞蛋白质合成的翻译后修饰的新方法和使用这种翻译后修饰反应的无细胞蛋白质合成的新方法。一种使用含有节肢动物衍生的提取物的无细胞蛋白质合成的无细胞蛋白质合成的无细胞系统中的蛋白质的合成方法，其特征在于，在哺乳动物来源的微粒体膜的存在下进行翻译反应。

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