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公开(公告)号:US20120100530A1
公开(公告)日:2012-04-26
申请号:US13147171
申请日:2010-01-29
申请人: Ruth Moysey , Michael Knaggs , Lakmal Jayasinghe , James White , Brian Mckeown , John Milton
发明人: Ruth Moysey , Michael Knaggs , Lakmal Jayasinghe , James White , Brian Mckeown , John Milton
CPC分类号: C12N9/22 , C07K14/31 , C07K2319/03 , C12N9/1241 , C12N9/90
摘要: The invention relates to constructs comprising a nucleic acid binding protein and a surface. At least one native accessible cysteine residue is removed from the binding protein. The binding protein is attached to the surface via one or more accessible cysteine residues. The removal of other accessible cysteine residues from the protein allows control attachment to the surface. The constructs can be used to generate transmembrane pores having a nucleic acid binding protein attached thereto. Such pores are particularly useful for sequencing nucleic acids. The enzyme handles the nucleic acid in such a way that the pore can detect each of its component nucleotides by stochastic sensing.
摘要翻译: 本发明涉及包含核酸结合蛋白和表面的构建体。 从结合蛋白中去除至少一种天然存在的半胱氨酸残基。 结合蛋白通过一个或多个可接近的半胱氨酸残基连接到表面。 从蛋白质中去除其他可获得的半胱氨酸残基允许控制附着于表面。 构建体可用于产生具有与其连接的核酸结合蛋白的跨膜孔。 这样的孔特别可用于测序核酸。 酶以这样的方式处理核酸,使得孔可以通过随机感测来检测其每个组分核苷酸。
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公开(公告)号:US20120064599A1
公开(公告)日:2012-03-15
申请号:US13147176
申请日:2010-01-29
申请人: Lakmal Jayasinghe , John Milton , Luke McNeill , James Clarke , James White , Ruth Moysey
发明人: Lakmal Jayasinghe , John Milton , Luke McNeill , James Clarke , James White , Ruth Moysey
CPC分类号: C12N9/22 , C07K14/31 , C07K2319/03 , C12N9/1241 , C12N9/90
摘要: The invention provides method of covalently coupling two or more moieties, the method comprising: (a) providing a first moiety having covalently attached thereto (i) at least one first linker comprising a first hybridizable region and (ii) at least one first group capable of forming a covalent bond; (b) providing a second moiety having covalently attached thereto (i) at least one second linker comprising a second hybridizable region capable of hybridizing to the first hybridizable region and (ii) at least a second group capable of forming a covalent bond with the first group; (c) contacting the first and second moieties under conditions that allow the first and second hybridizable regions to hybridize and link the moieties; and (d) exposing the linked moieties to conditions that allow the formation of a covalent bond between the first and second groups.
摘要翻译: 本发明提供了共价偶联两个或更多个部分的方法,所述方法包括:(a)提供共价连接到其上的第一部分(i)至少一个包含第一可杂交区域的第一接头和(ii)至少一个能够 形成共价键; (b)提供共价连接到其上的第二部分(i)至少一个第二接头,其包含能够与第一可杂交区域杂交的第二可杂交区域和(ii)至少第二个能够与第一可共< 组; (c)在允许第一和第二可杂交区域杂交并连接部分的条件下使第一和第二部分接触; 和(d)将连接的部分暴露于允许在第一和第二基团之间形成共价键的条件。
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公开(公告)号:US09222082B2
公开(公告)日:2015-12-29
申请号:US13147176
申请日:2010-01-29
申请人: Lakmal Jayasinghe , John Milton , Luke McNeill , James Anthony Clarke , James White , Ruth Moysey
发明人: Lakmal Jayasinghe , John Milton , Luke McNeill , James Anthony Clarke , James White , Ruth Moysey
CPC分类号: C12N9/22 , C07K14/31 , C07K2319/03 , C12N9/1241 , C12N9/90
摘要: The invention provides method of covalently coupling two or more moieties, the method comprising: (a) providing a first moiety having covalently attached thereto (i) at least one first linker comprising a first hybridizable region and (ii) at least one first group capable of forming a covalent bond; (b) providing a second moiety having covalently attached thereto (i) at least one second linker comprising a second hybridizable region capable of hybridizing to the first hybridizable region and (ii) at least a second group capable of forming a covalent bond with the first group; (c) contacting the first and second moieties under conditions that allow the first and second hybridizable regions to hybridize and link the moieties; and (d) exposing the linked moieties to conditions that allow the formation of a covalent bond between the first and second groups.
摘要翻译: 本发明提供了共价偶联两个或更多个部分的方法,所述方法包括:(a)提供共价连接到其上的第一部分(i)至少一个包含第一可杂交区域的第一接头和(ii)至少一个能够 形成共价键; (b)提供共价连接到其上的第二部分(i)至少一个第二接头,其包含能够与第一可杂交区域杂交的第二可杂交区域和(ii)至少第二个能够与第一可共< 组; (c)在允许第一和第二可杂交区域杂交并连接部分的条件下使第一和第二部分接触; 和(d)将连接的部分暴露于允许在第一和第二基团之间形成共价键的条件。
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公开(公告)号:US20110229877A1
公开(公告)日:2011-09-22
申请号:US13002709
申请日:2009-07-06
申请人: Lakmal Jayasinghe , Hagan Bayley , Stephen Cheley , Brian Mckeown , James White , James Clarke
发明人: Lakmal Jayasinghe , Hagan Bayley , Stephen Cheley , Brian Mckeown , James White , James Clarke
CPC分类号: C12Q1/6869 , C07K14/31 , C12N9/1247 , C12N9/1252 , C12N9/127 , C12N9/1276 , C12N9/16 , C12N9/22 , C12N9/52 , C12N9/90 , C12N9/96 , C12Q2565/631
摘要: The invention relates to constructs comprising a transmembrane protein pore subunit and a nucleic acid handling enzyme. The pore subunit is covalently attached to the enzyme such that both the subunit and enzyme retain their activity. The constructs can be used to generate transmembrane protein pores having a nucleic acid handling enzyme attached thereto. Such pores are particularly useful for sequencing nucleic acids. The enzyme handles the nucleic acid in such a way that the pore can detect its component nucleotides by stochastic sensing.
摘要翻译: 本发明涉及包含跨膜蛋白质孔亚基和核酸处理酶的构建体。 孔亚基共价连接到酶上,使得亚基和酶保持其活性。 构建体可用于产生具有附着于其上的核酸处理酶的跨膜蛋白孔。 这样的孔特别可用于测序核酸。 酶以这样的方式处理核酸,使得孔可以通过随机感测来检测其成分核苷酸。
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公开(公告)号:US09751915B2
公开(公告)日:2017-09-05
申请号:US13984628
申请日:2012-02-10
CPC分类号: C07K14/35 , A61K39/00 , A61K39/04 , C12Q1/6869
摘要: The invention relates to mutant forms of Msp. The invention also relates to nucleic acid characterization using Msp.
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公开(公告)号:US20140186823A1
公开(公告)日:2014-07-03
申请号:US13984628
申请日:2012-02-10
CPC分类号: C07K14/35 , A61K39/00 , A61K39/04 , C12Q1/6869
摘要: The invention relates to mutant forms of Msp. The invention also relates to nucleic acid characterisation using Msp.
摘要翻译: 本发明涉及Msp的突变形式。 本发明还涉及使用Msp的核酸表征。
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公开(公告)号:US09447152B2
公开(公告)日:2016-09-20
申请号:US13002717
申请日:2009-07-06
IPC分类号: C12Q1/68 , C07K14/245
CPC分类号: C07K14/245 , C12Q1/6869 , C12Q2565/631
摘要: The invention relates to a mutant α-hemolysin (α-HL) pore which is useful for detecting one or more nucleotides by stochastic sensing. The pore is particularly useful for sequencing DNA or RNA. A molecular adaptor that allows detection of the nucleotide(s) is covalently attached to the pore. The pore is specifically modified to facilitate positioning of the adaptor and may be modified to facilitate covalent attachment.
摘要翻译: 本发明涉及用于通过随机感测检测一个或多个核苷酸的突变α-溶血素(α-HL)孔。 该孔特别适用于测序DNA或RNA。 允许检测核苷酸的分子衔接子共价连接到孔。 孔被特别地修改以便于适配器的定位并且可以被修改以促进共价附着。
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公开(公告)号:US20110177498A1
公开(公告)日:2011-07-21
申请号:US13002717
申请日:2009-07-06
申请人: James Clarke , Lakmal Jayasinghe , Terence Reid , Hagan Bayley
发明人: James Clarke , Lakmal Jayasinghe , Terence Reid , Hagan Bayley
IPC分类号: C12Q1/68 , C07K14/195 , C07H21/04 , C12P21/00
CPC分类号: C07K14/245 , C12Q1/6869 , C12Q2565/631
摘要: The invention relates to a mutant α-hemolysin (α-HL) pore which is useful for detecting one or more nucleotides by stochastic sensing. The pore is particularly useful for sequencing DNA or RNA. A molecular adaptor that allows detection of the nucleotide(s) is covalently attached to the pore. The pore is specifically modified to facilitate positioning of the adaptor and may be modified to facilitate covalent attachment.
摘要翻译: 本发明涉及用于通过随机感测检测一个或多个核苷酸的突变α-溶血素(α-HL)孔。 该孔特别适用于测序DNA或RNA。 允许检测核苷酸的分子衔接子共价连接到孔。 孔被特别地修改以便于适配器的定位并且可以被修改以促进共价附着。
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公开(公告)号:US20180148481A2
公开(公告)日:2018-05-31
申请号:US15507947
申请日:2015-09-01
申请人: Stefan Howorka , Han Remaut , Lakmal Jayasinghe , Elizabeth Jayne Wallace , James Anthony Clarke , Richard George Hambley , Jonathan Bankes Pugh
发明人: Stefan Howorka , Han Remaut , Lakmal Jayasinghe , Elizabeth Jayne Wallace , James Anthony Clarke , Richard George Hambley , Jonathan Bankes Pugh
IPC分类号: C07K14/245 , C12Q1/68 , G01N27/447
CPC分类号: C07K14/245 , C07K2319/22 , C12Q1/68 , C12Q1/6869 , C12Q1/6874 , C12Q2565/631 , G01N27/447
摘要: The invention relates to mutant forms of CsgG. The invention also relates to analyte detection and characterisation using CsgG.
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