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1. 发明申请

US20120238470A1 METHOD FOR SCREENING AND QUANTIFYING VARIOUS ENZYME ACTIVITIES USING A GENETIC ENZYME SCREENING SYSTEM 有权
标题翻译：使用遗传酶筛选系统筛选和定量各种酶活性的方法
公开(公告)号：US20120238470A1
公开(公告)日：2012-09-20
申请号：US13376783
申请日：2010-06-08
申请人： Seung Goo Lee , Eugene Rha , Su Lim Choi , Jae Jun Song , Jong Hyun Choi , Hee Sik Kim
发明人： Seung Goo Lee , Eugene Rha , Su Lim Choi , Jae Jun Song , Jong Hyun Choi , Hee Sik Kim
IPC分类号： C40B30/08
CPC分类号： C12Q1/6897 , C12N15/1086
摘要： A method of detecting and quantifying various enzymatic activities using a constructed artificial genetic circuit GESS (genetic enzyme screening system) for sensing phenolic compounds and a method of screening a trace of activities of target enzymes from a metagenome using the artificial genetic circuit, thereby securing target enzyme genes. When the method for screening and quantifying target enzymatic activity is used, useful genes can be screened from various genetic communities, including environmental or metagenomic libraries, at a single cell level in high throughput (million/day). Further, the sensitivity of the genetic circuit to phenol derivatives and the expression thereof can be controlled, and thus the genetic circuit can rapidly sense and quantify various enzymatic activities. Thus, the method can be advantageously used in the protein engineering technology for enzyme modification. Particularly, it can quantitatively investigate enzymatic activity, and thus can be applied to molecular evolution technology.
摘要翻译：使用用于感测酚类化合物的构建的人工遗传回路GESS（遗传酶筛选系统）检测和定量各种酶活性的方法，以及使用人造遗传回路从宏基因组筛选靶酶的活性痕迹的方法，由此确保靶酶基因。当使用筛选和定量靶酶活性的方法时，可以在高通量（百万/天）的单个细胞水平上从各种遗传群落（包括环境或宏基因组文库）筛选有用的基因。此外，可以控制遗传回路对苯酚衍生物的敏感性及其表达，因此遗传回路可以快速感测和定量各种酶活性。因此，该方法可有利地用于酶修饰的蛋白质工程技术中。特别是可以定量研究酶活性，从而可以应用于分子进化技术。

2. 发明授权

US09783859B2 Method of screening and quantifying various enzymatic activities using artificial genetic circuits 有权
公开(公告)号：US09783859B2
公开(公告)日：2017-10-10
申请号：US13376783
申请日：2010-06-08
申请人： Seung Goo Lee , Eugene Rha , Su Lim Choi , Jae Jun Song , Jong Hyun Choi , Hee Sik Kim
发明人： Seung Goo Lee , Eugene Rha , Su Lim Choi , Jae Jun Song , Jong Hyun Choi , Hee Sik Kim
IPC分类号： C40B30/00 , C12Q1/68 , C12N15/10
CPC分类号： C12Q1/6897 , C12N15/1086
摘要： A method of detecting and quantifying various enzymatic activities using a constructed artificial genetic circuit GESS (genetic enzyme screening system) for sensing phenolic compounds and a method of screening a trace of activities of target enzymes from a metagenome using the artificial genetic circuit, thereby securing target enzyme genes. When the method for screening and quantifying target enzymatic activity is used, useful genes can be screened from various genetic communities, including environmental or metagenomic libraries, at a single cell level in high throughput (million/day). Further, the sensitivity of the genetic circuit to phenol derivatives and the expression thereof can be controlled, and thus the genetic circuit can rapidly sense and quantify various enzymatic activities. Thus, the method can be advantageously used in the protein engineering technology for enzyme modification. Particularly, it can quantitatively investigate enzymatic activity, and thus can be applied to molecular evolution technology.

3. 发明申请

US20130040366A1 METAGENOME-DERIVED ALKALINE PHOSPHATASE 有权
标题翻译： METAGENOME衍生的碱性磷酸酶
公开(公告)号：US20130040366A1
公开(公告)日：2013-02-14
申请号：US13519013
申请日：2010-06-08
申请人： Seung Goo Lee , Su Lim Choi , Eugene Rha , Jae Jun Song
发明人： Seung Goo Lee , Su Lim Choi , Eugene Rha , Jae Jun Song
IPC分类号： C12N9/16 , C12N1/15 , C12N1/21 , C12N1/19 , C12N15/55 , C12N15/63
CPC分类号： C12N9/16
摘要： The present invention relates to a metagenome-derived alkaline phosphatase, and more particularly to a novel, metagenome-derived alkaline phosphatase screened using a artificial genetic circuit that detects phenolic compounds, and a preparation method thereof. A novel alkaline phosphatase according to the present invention has high activity of dephosphorylating DNA and can be inactivated rapidly by simple heat treatment. Thus, it can be used for a dephosphorylation reaction so that genetic manipulations, including genetic cloning, become efficient. In addition, it can be actively expressed in recombinant microorganisms, and thus can be used in various assays, including enzyme immunoassay.
摘要翻译：本发明涉及一种来源于碱金属蛋白酶的碱性磷酸酶，更具体地说，涉及使用检测酚类化合物的人工遗传回路筛选的新颖的，基因突变型的碱性磷酸酶及其制备方法。根据本发明的新型碱性磷酸酶具有高的去磷酸化DNA的活性，并且可以通过简单的热处理快速灭活。因此，它可以用于去磷酸化反应，使得遗传操作（包括遗传克隆）变得有效。此外，它可以在重组微生物中有效表达，因此可用于各种测定，包括酶免疫测定。

4. 发明授权

US08647854B2 Metagenome-derived alkaline phosphatase 有权
标题翻译：来自Metagenome的碱性磷酸酶
公开(公告)号：US08647854B2
公开(公告)日：2014-02-11
申请号：US13519013
申请日：2010-06-08
申请人： Seung Goo Lee , Su Lim Choi , Eugene Rha , Jae Jun Song
发明人： Seung Goo Lee , Su Lim Choi , Eugene Rha , Jae Jun Song
IPC分类号： C12N9/16
CPC分类号： C12N9/16
摘要： The present invention relates to a metagenome-derived alkaline phosphatase, and more particularly to a novel, metagenome-derived alkaline phosphatase screened using a artificial genetic circuit that detects phenolic compounds, and a preparation method thereof. A novel alkaline phosphatase according to the present invention has high activity of dephosphorylating DNA and can be inactivated rapidly by simple heat treatment. Thus, it can be used for a dephosphorylation reaction so that genetic manipulations, including genetic cloning, become efficient. In addition, it can be actively expressed in recombinant microorganisms, and thus can be used in various assays, including enzyme immunoassay.
摘要翻译：本发明涉及一种来源于碱金属蛋白酶的碱性磷酸酶，更具体地说，涉及使用检测酚类化合物的人工遗传回路筛选的新颖的，基因突变型的碱性磷酸酶及其制备方法。根据本发明的新型碱性磷酸酶具有高的去磷酸化DNA的活性，并且可以通过简单的热处理快速灭活。因此，它可以用于去磷酸化反应，使得遗传操作（包括遗传克隆）变得有效。此外，它可以在重组微生物中有效表达，因此可用于各种测定，包括酶免疫测定。

5. 发明授权

US08877446B2 Method for detecting protein-protein interactions in cells 有权
标题翻译：检测细胞中蛋白质 - 蛋白质相互作用的方法
公开(公告)号：US08877446B2
公开(公告)日：2014-11-04
申请号：US13524868
申请日：2012-06-15
申请人： Seung Goo Lee , Su-Lim Choi , Jong Sik Gam , Jae Jun Song , Sang Jun Lee
发明人： Seung Goo Lee , Su-Lim Choi , Jong Sik Gam , Jae Jun Song , Sang Jun Lee
IPC分类号： C12Q1/68 , G01N33/542 , G01N33/50 , G01N21/64
CPC分类号： G01N21/6428 , G01N33/5008 , G01N33/542
摘要： The present invention relates to a method for detecting protein-protein interactions in living cells, and more particularly, to a method for providing cells comprising a first construct and a second construct, wherein the first construct comprises a polynucleotide encoding a first fusion protein which comprises a bait protein, a first fluorescent protein and a CBD (cellulose-binding domain) protein, and wherein the second construct comprises a polynucleotide encoding a second fusion protein which comprises a prey protein and a second fluorescent protein so as to allow formation of inclusion bodies, and detecting interactions between the bait protein and the prey protein that are displayed by inclusion bodies, a method for isolating the prey protein bound to the bait protein using the cells comprising the constructs, the cells, and a kit for detecting protein-protein interactions, comprising the constructs.
摘要翻译：本发明涉及一种用于检测活细胞中蛋白质 - 蛋白质相互作用的方法，更具体地说，涉及提供包含第一构建体和第二构建体的细胞的方法，其中第一构建体包含编码第一融合蛋白的多核苷酸，其包含诱饵蛋白质，第一荧光蛋白质和CBD（纤维素结合结构域）蛋白质，并且其中第二构建体包含编码第二融合蛋白的多核苷酸，其包含猎物蛋白质和第二荧光蛋白质，以便形成包涵体并检测由包涵体显示的诱饵蛋白和猎物蛋白之间的相互作用，使用包含构建体的细胞分离与诱饵蛋白结合的猎物蛋白的方法，所述细胞和用于检测蛋白质 - 蛋白质相互作用的试剂盒，包括构建体。

6. 发明申请

US20130052660A1 METHOD FOR DETECTING PROTEIN-PROTEIN INTERACTIONS IN CELLS 有权
标题翻译：检测细胞中蛋白质 - 蛋白质相互作用的方法
公开(公告)号：US20130052660A1
公开(公告)日：2013-02-28
申请号：US13524868
申请日：2012-06-15
申请人： Seung Goo Lee , Su-Lim Choi , Jong Sik Gam , Jae Jun Song , Sang Jun Lee
发明人： Seung Goo Lee , Su-Lim Choi , Jong Sik Gam , Jae Jun Song , Sang Jun Lee
IPC分类号： C12P21/02 , C12N1/21 , C12N5/10 , G01N21/64
CPC分类号： G01N21/6428 , G01N33/5008 , G01N33/542
摘要： The present invention relates to a method for detecting protein-protein interactions in living cells, and more particularly, to a method for providing cells comprising a first construct and a second construct, wherein the first construct comprises a polynucleotide encoding a first fusion protein which comprises a bait protein, a first fluorescent protein and a CBD (cellulose-binding domain) protein, and wherein the second construct comprises a polynucleotide encoding a second fusion protein which comprises a prey protein and a second fluorescent protein so as to allow formation of inclusion bodies, and detecting interactions between the bait protein and the prey protein that are displayed by inclusion bodies, a method for isolating the prey protein bound to the bait protein using the cells comprising the constructs, the cells, and a kit for detecting protein-protein interactions, comprising the constructs.
摘要翻译：本发明涉及一种用于检测活细胞中蛋白质 - 蛋白质相互作用的方法，更具体地说，涉及提供包含第一构建体和第二构建体的细胞的方法，其中第一构建体包含编码第一融合蛋白的多核苷酸，其包含诱饵蛋白质，第一荧光蛋白质和CBD（纤维素结合结构域）蛋白质，并且其中第二构建体包含编码第二融合蛋白的多核苷酸，其包含猎物蛋白质和第二荧光蛋白质，以便形成包涵体并检测由包涵体显示的诱饵蛋白和猎物蛋白之间的相互作用，使用包含构建体的细胞分离与诱饵蛋白结合的猎物蛋白的方法，所述细胞和用于检测蛋白质 - 蛋白质相互作用的试剂盒，包括构建体。

7. 发明授权

US09921215B2 Method for detecting ligand using FRET biosensor 有权
公开(公告)号：US09921215B2
公开(公告)日：2018-03-20
申请号：US13266753
申请日：2010-04-27
申请人： Seung Goo Lee , Jae Seok Ha , Jae Jun Song
发明人： Seung Goo Lee , Jae Seok Ha , Jae Jun Song
IPC分类号： G01N33/542 , B82Y15/00 , G01N33/58
CPC分类号： G01N33/542 , B82Y15/00 , G01N33/582 , G01N33/587 , G01N33/588 , G01N2500/00
摘要： The present application relates to a method for detecting ligand using a biosensor applied the FRET (fluorescence resonance energy transfer) phenomenon. More particularly, the method may be used for simply detecting a ligand in a sample by measuring the FRET of a biosensor under the conditions in which a specific critical temperature is maintained. The method may use a phenomenon in which a ligand-binding protein in a biosensor shows reversible unfolding at a temperature higher than the specific critical temperature and the level of the unfolding changes depending on the concentration of a ligand. The method can be widely applied to a variety of kinds of FRET biosensors using the ligand-binding protein.

8. 发明授权

US07323328B2 Obligately symbiotic thermophile Symbiobacterium toebii SC-1 producing thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase 有权
标题翻译：共生共生嗜热菌共培养杆菌SC-1产生热稳定性L-酪氨酸苯酚裂解酶和L-色氨酸吲哚裂解酶
公开(公告)号：US07323328B2
公开(公告)日：2008-01-29
申请号：US11186559
申请日：2005-07-21
申请人： Moon-Hee Sung , Seung-Goo Lee , Sung-Keun Rhee , Seung-Pyo Hong , Dae-Joong Kang , Su-Mi Kim , Chul-Joong Kim , Jin-Woo Bae , Che Ok Jeon , Joong-Jae Kim , Kwang Kim , Jae Jun Song
发明人： Moon-Hee Sung , Seung-Goo Lee , Sung-Keun Rhee , Seung-Pyo Hong , Dae-Joong Kang , Su-Mi Kim , Chul-Joong Kim , Jin-Woo Bae , Che Ok Jeon , Joong-Jae Kim , Kwang Kim , Jae Jun Song
IPC分类号： C12N1/20 , C12N9/88 , C12P39/00
CPC分类号： C12R1/01 , C12N9/88 , C12P13/227
摘要： The present invention relates to a novel obligately symbiotic thermophile Symbiobacterium toebii SC-1 (Accession NO: KCTC 0685BP), and a thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1. The present invention also relates to a pure culture method of Symbiobacterium toebii SC-1, a method of growth measurement for Symbiobacterium toebii SC-1 and its relative symbiotic bacteria showing low-growth yield using nitrate respiration, and a screening method of relative symbiotic strains of Symbiobacterium toebii SC-1 from the environment using a specific antibody to it. Therefore, the thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1 in this invention can provide more stable catalysts in enzymatic biotransformation processes which enzymatically produce valuable medico-amino acids 3,4-dehydroxy-L-phenylalanine (L-DOPA) and L-tryptophan.
摘要翻译：本发明涉及一种新型的共生共生嗜热ile ile i i i i i i i i））i i i i i i i i i i i i i i i i i。。。。。。。。。。。。。。。。。。。。本发明还涉及使用硝酸盐呼吸的共生杆菌SC-1的纯培养方法，使用硝酸盐呼吸显示低生长产量的共生杆菌SC-1及其相对共生菌的生长方法，以及相对共生菌株的筛选方法的来自环境的共表达杆菌SC-1使用特异性抗体。因此，在本发明中由共生杆菌（Toibii）SC-1产生的热稳定性L-酪氨酸苯酚裂解酶和L-色氨酸吲哚裂解酶可以在酶生物转化过程中提供更稳定的催化剂，其中酶促产生有价值的羟基 - 氨基酸3,4-脱羟基 - L-苯丙氨酸（L-DOPA）和L-色氨酸。

9. 发明申请

US20120083048A1 METHOD FOR DETECTING LIGAND USING FRET BIOSENSOR 有权
标题翻译：使用FRET生物传感器检测配体的方法
公开(公告)号：US20120083048A1
公开(公告)日：2012-04-05
申请号：US13266753
申请日：2010-04-27
申请人： Seung Goo Lee , Jae Seok Ha , Jae Jun Song
发明人： Seung Goo Lee , Jae Seok Ha , Jae Jun Song
IPC分类号： G01N33/566 , B82Y15/00
CPC分类号： G01N33/542 , B82Y15/00 , G01N33/582 , G01N33/587 , G01N33/588 , G01N2500/00
摘要： The present application relates to a method for detecting ligand using a biosensor applied the FRET(fluorescence resonance energy transfer) phenomenon. More particularly, the method may be used for simply detecting a ligand in a sample by measuring the FRET of a biosensor under the conditions in which a specific critical temperature is maintained. The method may use a phenomenon in which a ligand-binding protein in a biosensor shows reversible unfolding at a temperature higher than the specific critical temperature and the level of the unfolding changes depending on the concentration of a ligand. The method can be widely applied to a variety of kinds of FRET biosensors using the ligand-binding protein.
摘要翻译：本申请涉及使用应用FRET（荧光共振能量转移）现象的生物传感器检测配体的方法。更具体地说，该方法可用于通过在保持特定临界温度的条件下测量生物传感器的FRET来简单地检测样品中的配体。该方法可以使用生物传感器中的配体结合蛋白在比特定临界温度高的温度下显示可逆解折叠的现象，并且解折叠水平根据配体的浓度而变化。该方法可广泛应用于使用配体结合蛋白的各种FRET生物传感器。

10. 发明授权

US06967087B1 Obligately symbiotic thermophile symbiobacterium toebii SC-1 producing thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase and a method for screening its relative bacteria 失效
标题翻译：共生共生嗜热菌共生菌toebii SC-1产生热稳定性L-酪氨酸苯酚裂解酶和L-色氨酸吲哚裂解酶及其相对细菌筛选方法
公开(公告)号：US06967087B1
公开(公告)日：2005-11-22
申请号：US10220174
申请日：2000-07-14
申请人： Moon-Hee Sung , Seung-Goo Lee , Sung-Keun Rhee , Seung-Pyo Hong , Dae-Joong Kang , Su-Mi Kim , Chul-Joong Kim , Jin-Woo Bae , Che Ok Jeon , Joong-Jae Kim , Kwang Kim , Jae Jun Song
发明人： Moon-Hee Sung , Seung-Goo Lee , Sung-Keun Rhee , Seung-Pyo Hong , Dae-Joong Kang , Su-Mi Kim , Chul-Joong Kim , Jin-Woo Bae , Che Ok Jeon , Joong-Jae Kim , Kwang Kim , Jae Jun Song
IPC分类号： G01N33/569 , C12N1/20 , C12N9/88 , C12P13/22 , C12Q1/06 , C12R1/01 , G01N21/77 , C12P39/00 , C12Q1/12
CPC分类号： C12R1/01 , C12N9/88 , C12P13/227
摘要： The present invention relates to a novel obligately symbiotic thermophile Symbiobacterium toebii SC-1(Accession NO: KCTC 0685BP), and a thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1. The present invention also relates to a pure culture method of Symbiobacterium toebii SC-1, a method of growth measurement for Symbiobacterium toebu SC-1 and its relative symbiotic bacteria showing low-growth yield using nitrate respiration, and a screening method of relative symbiotic strains of Symbiobacterium toebii SC-1 from the environment using a specific antibody to it. Therefore, the thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase produced by Symbiobacterium toebii SC-1 in this invention can provide more stable catalysts in enzymatic biotransformation processes which enzymatically produce valuable medico-amino acids 3,4-dehydroxy-L-phenylalanine (L-DOPA) and L-tryptophan.
摘要翻译：本发明涉及一种新型的共生共生嗜热ile ile i i i i i i i i））i i i i i i i i i i i i i i i i i。。。。。。。。。。。。。。。。。。。。本发明还涉及使用硝酸盐呼吸的共生杆菌SC-1的纯培养方法，使用硝酸盐呼吸显示低生长量的共生杆菌SC-1及其相对共生菌的生长测定方法以及相对共生菌株的筛选方法的来自环境的共表达杆菌SC-1使用特异性抗体。因此，在本发明中由共生杆菌（Toibii）SC-1产生的热稳定性L-酪氨酸苯酚裂解酶和L-色氨酸吲哚裂解酶可以在酶生物转化过程中提供更稳定的催化剂，其中酶促产生有价值的羟基 - 氨基酸3,4-脱羟基 - L-苯丙氨酸（L-DOPA）和L-色氨酸。

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