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1. 发明申请

US20110244521A1 METHOD FOR PREPARATION OF RECOBINANT DNA 审中-公开
标题翻译：回收DNA的制备方法
公开(公告)号：US20110244521A1
公开(公告)日：2011-10-06
申请号：US12674160
申请日：2008-08-12
申请人： Takeharu Nagai , Ippei Kotera
发明人： Takeharu Nagai , Ippei Kotera
IPC分类号： C12P19/34
CPC分类号： C12N15/64 , C12N15/66
摘要： The problem to be solved in the present invention is to provide a simplified and efficiently improved DNA recombination method.The above problem can be solved with the present method for preparing a recombinant DNA by inserting a DNA fragment of interest into a vector DNA, the method comprising the step of carrying out the following reactions at the same reacting location at the substantially simultaneouse time: (a) a reaction for simultaneously cleaving a site of the vector for inserting the fragment and a DNA containing the fragment in the presence of a restriction enzyme whose DNA recognition site and DNA cleavage site are discrete; and (b) a reaction for inserting the fragment into the vector in the presence of a DNA ligase.
摘要翻译：本发明要解决的问题是提供简化且有效改进的DNA重组方法。通过将目的DNA片段插入载体DNA中制备重组DNA的本发明的方法可以解决上述问题，该方法包括在大致同步的时间在相同的反应位置进行以下反应的步骤：（ a）在DNA识别位点和DNA切割位点离散的限制性酶的存在下，同时切割用于插入片段的载体的位点和含有该片段的DNA的反应; 和（b）在DNA连接酶存在下将片段插入载体的反应。

2. 发明授权

US07981658B2 Nucleic acid molecules, vectors, and host cells encoding fluorescent protein 有权
标题翻译：核酸分子，载体和编码荧光蛋白的宿主细胞
公开(公告)号：US07981658B2
公开(公告)日：2011-07-19
申请号：US12569464
申请日：2009-09-29
申请人： Atsushi Miyawaki , Takeharu Nagai
发明人： Atsushi Miyawaki , Takeharu Nagai
IPC分类号： C12N1/20 , C12N15/00 , C07K1/00 , C07H21/04
CPC分类号： C07K14/43595 , C07K2319/00
摘要： The present invention provides a fluorescent protein having an amino acid sequence of the green fluorescent protein, the yellow fluorescent protein or mutants thereof wherein the 46th phenylalanine residue is substituted with a leucine residue. According to the present invention, there are provided novel GFP or YEP mutants having an excellent maturation efficacy and having a decreased sensitivity to both of H+ and CI31.
摘要翻译：本发明提供了具有绿色荧光蛋白的氨基酸序列，黄色荧光蛋白或其突变体的荧光蛋白，其中第46位苯丙氨酸残基被亮氨酸残基取代。根据本发明，提供了具有优异成熟功效并且对H +和CI31都具有降低的敏感性的新型GFP或YEP突变体。

3. 发明申请

US20100009354A1 METHOD FOR DETERMINING BASE SEQUENCE OF DNA 失效
标题翻译：确定DNA基因序列的方法
公开(公告)号：US20100009354A1
公开(公告)日：2010-01-14
申请号：US12279924
申请日：2007-02-20
申请人： Takeharu Nagai , Tomomi Tani , Ippei Kotera , Yoshihiro Yoneda
发明人： Takeharu Nagai , Tomomi Tani , Ippei Kotera , Yoshihiro Yoneda
IPC分类号： C12Q1/68 , C07H21/04 , C12M1/34
CPC分类号： C12Q1/6869 , C12Q2563/107 , C12Q2521/313 , C12Q2521/501
摘要： The present invention provides a method for determining the base sequence of a DNA. According to the method for determining the base sequence of a DNA of the present invention, a probe is used, which is a probe having a protruding end and identification-labeled according to the species of the base at the protruding end, containing a recognition sequence of a class IIS restriction enzyme, to carry out simultaneously in a chain reaction, for a plurality of DNAs to be analyzed, ligation of the end base of a DNA to be analyzed and a probe and cleavage of the end base of the DNA to be analyzed, allowing the base sequence to be determined sequentially by a single molecule spectrofluorimetry method, such that an effective determination of the base sequence of a DNA becomes possible.
摘要翻译：本发明提供了确定DNA碱基序列的方法。根据本发明的DNA的碱基序列的测定方法，使用探针，该探针是具有突出端的探针，根据突起端的碱基的种类鉴定标记，其含有识别序列的IIS类限制酶，在连锁反应中同时进行多个要分析的DNA，连接待分析的DNA的末端碱基和探针，并将DNA的末端碱基切割成分析，通过单分子分光荧光法依次确定碱基序列，使得DNA的碱基序列的有效测定成为可能。

4. 发明授权

US07283306B2 Confocal optical scanner 失效
标题翻译：共焦光学扫描仪
公开(公告)号：US07283306B2
公开(公告)日：2007-10-16
申请号：US10844421
申请日：2004-05-13
申请人： Kenta Mikuriya , Takeo Tanaami , Naoki Seki , Takeharu Nagai , Atsushi Miyawaki
发明人： Kenta Mikuriya , Takeo Tanaami , Naoki Seki , Takeharu Nagai , Atsushi Miyawaki
IPC分类号： G02B27/14 , G02B21/00 , G02B26/02
CPC分类号： G02B21/0024 , G02B21/0044 , G02B21/0076
摘要： A confocal optical scanner of Nipkow disk type for measuring a sample, in which each component of two or more color pigments, has a high reflection mirror for separating an excited light to be radiated onto the sample and a fluorescence emitted from the sample, wherein a reflectance of the high reflection mirror is from 80% to 100% in a measured wavelength region including at least an excited light wavelength region and a fluorescence wavelength region. Thereby, the confocal optical scanner capable of measuring a polychromatic fluorescent image at the same time is realized with the enhanced light receiving efficiency of fluorescence, using one high reflection mirror without using a plurality of dichroic mirrors by exchange.
摘要翻译：用于测量样品的Nipkow盘类型的共焦光学扫描仪，其中两种或更多种颜色颜料的每种组分具有用于分离待辐射到样品上的激发光和从样品发射的荧光的高反射镜，其中a 在一个测量的波长区域中，至少包括激发光波长区域和荧光波长区域，高反射镜的反射率为80％至100％。由此，能够通过使用一个高反射镜而不使用多个二向色镜通过交换来实现能够同时测量多色荧光图像的共焦光学扫描仪，其具有增强的荧光的光接收效率。

5. 发明申请

US20060216075A1 Transfer device and image forming device 有权
标题翻译：转印装置和成像装置
公开(公告)号：US20060216075A1
公开(公告)日：2006-09-28
申请号：US11377429
申请日：2006-03-17
申请人： Hirokatsu Sugawara , Takeharu Nagai
发明人： Hirokatsu Sugawara , Takeharu Nagai
IPC分类号： G03G15/16
CPC分类号： G03G15/6558 , G03G15/1665 , G03G2215/00675
摘要： A transfer device for transferring a toner image formed on an image carrier onto a recording sheet includes a transfer assisting blade adapted to abut the recording sheet and press the recording sheet against the image carrier, the transfer assisting blade having a slant formed in a transverse direction of the blade at a front end of the blade which end is for abutment against the recording sheet, the slant having an angle relative to an advancing direction of the recording sheet.
摘要翻译：用于将形成在图像载体上的调色剂图像转印到记录纸上的转印装置包括转印辅助刮板，其适于邻接记录纸并将记录纸张压靠在图像载体上，转印辅助刮板具有沿横向形成的倾斜在叶片的前端处的叶片的端部用于抵靠记录片材，该倾斜件相对于记录片材的前进方向具有一定角度。

6. 发明申请

US20100167394A1 ULTRAMARINE FLUORESCENT PROTEIN 审中-公开
标题翻译：超紫外线荧光蛋白
公开(公告)号：US20100167394A1
公开(公告)日：2010-07-01
申请号：US12453000
申请日：2008-08-01
申请人： Takeharu Nagai , Wataru Tomosugi , Tomoki Matsuda
发明人： Takeharu Nagai , Wataru Tomosugi , Tomoki Matsuda
IPC分类号： C12N5/10 , C07K14/00 , C07H21/00 , C12N15/74
CPC分类号： C07K14/43595 , C07K2319/60
摘要： The present invention provides an artificial mutant of GFP having a novel emission peak, i.e., a fluorescent protein having an emission peak at 424 nm comprising an amino acid sequence represented by SEQ ID NO: 1, in which each of the amino acid residues at the 66th position and the 175th position is replaced and at least one of the amino acid residues at the 72nd position and the 206th position is further replaced, or a fluorescent protein having an emission peak at 424 nm and a pH-independent fluorescence intensity, in which each of the amino acid residues at the 65th, 145th, 148th, 46th and/or 203rd positions is further substituted. The fluorescent protein of the invention emits fluorescence having an emission peak at 424 nm and can be visually distinguished by its ultramarine color from other fluorescent proteins. The fluorescent protein has a pH-independent fluorescence intensity which is not affected by pH changes.
摘要翻译：本发明提供具有新发射峰的GFP的人工突变体，即在424nm处具有发射峰的荧光蛋白，其包含由SEQ ID NO：1表示的氨基酸序列，其中每个氨基酸残基第66位和第175位被置换，第72位和第206位的氨基酸残基中的至少一个被进一步置换，或者在424nm具有发射峰和不依赖于pH的荧光强度的荧光蛋白，其中第65，第145，第148，第46和/或第203位的每个氨基酸残基被进一步取代。本发明的荧光蛋白发射具有424nm处的发射峰的荧光，并且可以通过其来自其他荧光蛋白的群青颜色来目视辨别荧光。荧光蛋白具有pH不依赖于荧光强度，不受pH变化的影响。

7. 发明授权

US07595375B2 Fluorescent protein 有权
标题翻译：荧光蛋白
公开(公告)号：US07595375B2
公开(公告)日：2009-09-29
申请号：US10162593
申请日：2002-06-06
申请人： Atsushi Miyawaki , Takeharu Nagai
发明人： Atsushi Miyawaki , Takeharu Nagai
IPC分类号： C07K14/00 , C07H21/04 , C12N1/20 , C12N15/00
CPC分类号： C07K14/43595 , C07K2319/00
摘要： The present invention provides a fluorescent protein having an amino acid sequence of the green fluorescent protein, the yellow fluorescent protein or mutants thereof wherein the 46th phenylalanine residue is substituted with a leucine residue. According to the present invention, there are provided novel GFP or YEP mutants having an excellent maturation efficacy and having a decreased sensitivity to both of H+ and Cl−.
摘要翻译：本发明提供了具有绿色荧光蛋白的氨基酸序列，黄色荧光蛋白或其突变体的荧光蛋白，其中第46位苯丙氨酸残基被亮氨酸残基取代。根据本发明，提供了具有优异成熟功效并且对H +和Cl-两者具有降低的敏感性的新型GFP或YEP突变体。

8. 发明申请

US20090176211A1 FLUORESCENT PROTEIN 失效
标题翻译：荧光蛋白
公开(公告)号：US20090176211A1
公开(公告)日：2009-07-09
申请号：US11719166
申请日：2005-11-14
申请人： Takeharu Nagai , Atsushi Miyawaki
发明人： Takeharu Nagai , Atsushi Miyawaki
IPC分类号： C12Q1/68 , C07K14/00 , C12N15/11 , C12N15/00 , C12N5/06
CPC分类号： G01N33/5035 , C07K14/43595 , G01N33/542
摘要： It is an object of the present invention to provide a fluorescent protein, which allows an acceptor for fluorescence resonance energy transfer (FRET) to appear in a stimulating light-dependent manner, thereby enabling the marking of any given cell organelle, cells, or tissues, with multiple colors. The present invention provides a fluorescent protein which consists of a fused protein of a donor fluorescent protein and an acceptor fluorescent protein, wherein before irradiation with stimulating light, the donor protein is able to emit fluorescence as a result of irradiation of the donor protein with excitation light; and after irradiation with stimulating light, intramolecular FRET occurs between the donor fluorescent protein and the acceptor fluorescent protein as a result of irradiation of the donor protein with excitation light, and the acceptor protein is able to emit fluorescence, and wherein the fluorescence of the donor protein and the fluorescence of the acceptor protein have wavelengths that are different from each other.
摘要翻译：本发明的目的是提供一种荧光蛋白，其允许荧光共振能量转移受体（FRET）以刺激光依赖的方式出现，从而能够标记任何给定的细胞器，细胞或组织，具有多种颜色。本发明提供了由供体荧光蛋白的融合蛋白和受体荧光蛋白组成的荧光蛋白，其中在用刺激光照射之前，供体蛋白能够通过激发照射供体蛋白而发出荧光光; 并且在用刺激光照射后，作为供体蛋白质与激发光照射的结果，在供体荧光蛋白和受体荧光蛋白之间发生分子内FRET，并且受体蛋白能够发射荧光，并且其中供体的荧光蛋白质，受体蛋白质的荧光具有彼此不同的波长。

9. 发明申请

US20090017516A1 TARGET PHYSIOLOGICAL FUNCTION INACTIVATOR USING PHOTOSENSITIZER-LABELED FLUORESCENT PROTEIN 审中-公开
标题翻译：使用光敏剂标记荧光蛋白的目标生理功能失活剂
公开(公告)号：US20090017516A1
公开(公告)日：2009-01-15
申请号：US11722283
申请日：2005-12-21
申请人： Takeharu Nagai , Atsushi Miyawaki
发明人： Takeharu Nagai , Atsushi Miyawaki
IPC分类号： C12N13/00 , C07K14/00 , C07F19/00 , C07K1/00
CPC分类号： C07K14/43595 , A61K41/0057 , A61K41/0071
摘要： An object of the present invention is to provide a method of generating reactive oxygen species in a light irradiation-dependent manner, so as to inactivate any target physiological function. The present invention provides a target physiological function inactivator which consists of a photosensitizer-labeled fluorescent protein, wherein fluorescence resonance energy transfer (FRET) from the fluorescent protein to the photosensitizer occurs as a result of light irradiation, so that the photosensitizer can be excited to generate reactive oxygen species.
摘要翻译：本发明的目的是提供一种依照光依赖的方式产生活性氧的方法，从而使任何目标生理功能失活。本发明提供了由光敏剂标记的荧光蛋白组成的目标生理功能失活剂，其中由于光照射而发生从荧光蛋白到光敏剂的荧光共振能量转移（FRET），使得光敏剂可被激发产生活性氧。

10. 发明授权

US07424258B2 Transfer device and image forming device that include a transfer assisting blade having a slant portion for abutting a recording medium 有权
标题翻译：传送装置和图像形成装置，其包括具有用于邻接记录介质的倾斜部分的转移辅助刀片
公开(公告)号：US07424258B2
公开(公告)日：2008-09-09
申请号：US11377429
申请日：2006-03-17
申请人： Hirokatsu Sugawara , Takeharu Nagai
发明人： Hirokatsu Sugawara , Takeharu Nagai
IPC分类号： G03G15/16
CPC分类号： G03G15/6558 , G03G15/1665 , G03G2215/00675
摘要： A transfer device for transferring a toner image formed on an image carrier onto a recording sheet includes a transfer assisting blade adapted to abut the recording sheet and press the recording sheet against the image carrier, the transfer assisting blade having a slant formed in a transverse direction of the blade at a front end of the blade which end is for abutment against the recording sheet, the slant having an angle relative to an advancing direction of the recording sheet.
摘要翻译：用于将形成在图像载体上的调色剂图像转印到记录纸上的转印装置包括转印辅助刮板，其适于邻接记录纸并将记录纸张压靠在图像载体上，转印辅助刮板具有沿横向形成的倾斜在叶片的前端处的叶片的端部用于抵靠记录片材，该倾斜件相对于记录片材的前进方向具有一定角度。

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