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    MUTANT PROTEASE BIOSENSORS WITH ENHANCED DETECTION CHARACTERISTICS
    1.
    发明申请
    MUTANT PROTEASE BIOSENSORS WITH ENHANCED DETECTION CHARACTERISTICS 有权
    具有增强检测特性的突变型PROTEASE生物传感器

    公开(公告)号：US20110283373A1

    公开(公告)日：2011-11-17

    申请号：US13105648

    申请日：2011-05-11

    申请人： Brock BINKOWSKI ,  Braeden BUTLER ,  Lance P. ENCELL ,  Frank FAN ,  Brad HOOK ,  Paul OTTO ,  Gediminas VIDUGIRIS ,  Susan WIGDAL ,  Kristopher ZIMMERMAN

    发明人： Brock BINKOWSKI ,  Braeden BUTLER ,  Lance P. ENCELL ,  Frank FAN ,  Brad HOOK ,  Paul OTTO ,  Gediminas VIDUGIRIS ,  Susan WIGDAL ,  Kristopher ZIMMERMAN

    IPC分类号： A01K67/00 ,  C07H21/00 ,  C12N1/00 ,  C12N15/63 ,  C12N5/10

    CPC分类号： A61K49/0045 ,  A01K67/0275 ,  A61K49/0013 ,  C12N9/0069 ,  C12Q1/37 ,  C12Q1/66 ,  C12Y113/12007

    摘要： A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.

    摘要翻译： 编码包含修饰的环状置换的热稳定性荧光素酶的生物传感器多肽的多核苷酸和将热稳定性荧光素酶的C末端部分连接到热稳定性萤光素酶的N末端部分的接头。 相对于亲本循环置换的热稳定性荧光素酶，修饰的环状置换的热稳定性荧光素酶被修饰。 接头包含能够与细胞中的靶分子相互作用的传感器区域。 经修饰的环状置换的热稳定荧光素酶在目标分子存在下相对于亲本循环置换的热稳定性荧光素酶在生物传感器与靶分子相互作用后具有增强的响应。 或者，修饰的环状置换的热稳定荧光素酶在生物传感器与目标分子相互作用后相对于在不存在目标分子的情况下相对于经修饰的环状置换的热稳定性荧光素酶具有增强的响应。

    Mutant protease biosensors with enhanced detection characteristics
    2.
    发明授权
    Mutant protease biosensors with enhanced detection characteristics 有权
    具有增强检测特性的突变蛋白酶生物传感器

    公开(公告)号：US08735559B2

    公开(公告)日：2014-05-27

    申请号：US13105648

    申请日：2011-05-11

    申请人： Brock Binkowski ,  Braeden Butler ,  Lance P. Encell ,  Frank Fan ,  Brad Hook ,  Paul Otto ,  Gediminas Vidugiris ,  Susan Wigdal ,  Kristopher Zimmerman

    发明人： Brock Binkowski ,  Braeden Butler ,  Lance P. Encell ,  Frank Fan ,  Brad Hook ,  Paul Otto ,  Gediminas Vidugiris ,  Susan Wigdal ,  Kristopher Zimmerman

    IPC分类号： C12N15/62 ,  C12N15/63 ,  C12N15/54 ,  C12N15/74 ,  C12N15/79

    CPC分类号： A61K49/0045 ,  A01K67/0275 ,  A61K49/0013 ,  C12N9/0069 ,  C12Q1/37 ,  C12Q1/66 ,  C12Y113/12007

    摘要： A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.

    摘要翻译： 编码包含修饰的环状置换的热稳定性荧光素酶的生物传感器多肽的多核苷酸和将热稳定性荧光素酶的C末端部分连接到热稳定性萤光素酶的N末端部分的接头。 相对于亲本循环置换的热稳定性荧光素酶，修饰的环状置换的热稳定性荧光素酶被修饰。 接头包含能够与细胞中的靶分子相互作用的传感器区域。 经修饰的环状置换的热稳定荧光素酶在目标分子存在下相对于亲本循环置换的热稳定性荧光素酶在生物传感器与靶分子相互作用后具有增强的响应。 或者，修饰的环状置换的热稳定荧光素酶在生物传感器与目标分子相互作用后相对于在不存在目标分子的情况下相对于经修饰的环状置换的热稳定性荧光素酶具有增强的响应。

    Method of error reduction in nucleic acid populations
    3.
    发明申请
    Method of error reduction in nucleic acid populations 有权
    核酸群体误差减少的方法

    公开(公告)号：US20060127926A1

    公开(公告)日：2006-06-15

    申请号：US11214201

    申请日：2005-08-29

    申请人： Peter Belshaw ,  Michael Sussman ,  Franco Cerrina ,  James Kaysen ,  Brock Binkowski ,  Kathryn Richmond

    发明人： Peter Belshaw ,  Michael Sussman ,  Franco Cerrina ,  James Kaysen ,  Brock Binkowski ,  Kathryn Richmond

    IPC分类号： C12Q1/68 ,  C12P19/34

    CPC分类号： C12N15/1027 ,  C12Q1/686 ,  C12Q2537/113 ,  C12Q2521/514 ,  C12Q2521/301

    摘要： A method is disclosed for the direct synthesis of double stranded DNA molecules of a variety of sizes and with any desired sequence. The DNA molecule to be synthesis is logically broken up into smaller overlapping DNA segments. A maskless microarray synthesizer is used to make a DNA microarray on a substrate in which each element or feature of the array is populated by DNA of a one of the overlapping DNA segments. The complement of each segment is also made in the microarray. The DNA segments are released from the substrate and held under conditions favoring hybridization of DNA, under which conditions the segments will hybridize to form duplexes. The duplexes are then separated using a DNA binding agent which binds to improperly formed DNA helixes to remove errors from the set of DNA molecules. The segments can then be hybridized to each other to assemble the larger target DNA sequence.

    摘要翻译： 公开了用于直接合成各种尺寸和任何所需顺序的双链DNA分子的方法。 要合成的DNA分子在逻辑上分解成更小的重叠DNA片段。 无掩模微阵列合成器用于在基底上制备DNA微阵列，其中阵列的每个元件或特征由重叠DNA片段之一的DNA填充。 每个片段的补体也在微阵列中制成。 DNA片段从底物释放并保持在有利于DNA杂交的条件下，在该条件下片段将杂交形成双链体。 然后使用结合不正确形成的DNA螺旋的DNA结合剂分离双链体以从DNA分子组中除去错误。 然后可以将这些片段彼此杂交以组装较大的靶DNA序列。

    Permuted and nonpermuted luciferase biosensors
    4.
    发明授权
    Permuted and nonpermuted luciferase biosensors 有权
    有色和非精子萤光素酶生物传感器

    公开(公告)号：US09359635B2

    公开(公告)日：2016-06-07

    申请号：US11732105

    申请日：2007-04-02

    申请人： Brock Binkowski ,  Frank Fan ,  Susan Wigdal ,  Keith V. Wood ,  Monika G. Wood

    发明人： Brock Binkowski ,  Frank Fan ,  Susan Wigdal ,  Keith V. Wood ,  Monika G. Wood

    IPC分类号： C12N9/02 ,  C07H21/04 ,  C12N15/00 ,  C12N1/20 ,  C12N1/00 ,  C12Q1/66

    CPC分类号： C12N9/0069 ,  C12N9/0004 ,  C12Q1/66 ,  C12Y113/12007 ,  G01N33/84

    摘要： A modified luciferase protein which is a sensor for molecules including cAMP, cGMP, calcium, chelators thereof, kinases, or phosphatases is provided. Also provided is a circularly permuted anthozoan luciferase protein and a decapod crustacean luciferase protein, optionally containing one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest. Further provided is a modified anthozoan luciferase protein and a decapod crustacean luciferase protein containing an insertion of one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest.

    摘要翻译： 提供了一种修饰的荧光素酶蛋白，其是用于分子的传感器，包括cAMP，cGMP，钙，其螯合剂，激酶或磷酸酶。 还提供了循环置换的anthozoan萤光素酶蛋白质和十足配甲壳素荧光素酶蛋白质，任选地含有一个或多个异源氨基酸序列，其中至少一个直接或间接与感兴趣的分子相互作用。 进一步提供了一种修饰的anthozoan萤光素酶蛋白质和十足类甲壳类荧光素酶蛋白质，其含有一个或多个异源氨基酸序列的插入，其中至少一个与感兴趣的分子直接或间接相互作用。

    Permuted and nonpermuted luciferase biosensors
    6.
    发明申请
    Permuted and nonpermuted luciferase biosensors 有权
    有色和非精子萤光素酶生物传感器

    公开(公告)号：US20110039257A1

    公开(公告)日：2011-02-17

    申请号：US11732105

    申请日：2007-04-02

    申请人： Brock Binkowski ,  Frank Fan ,  Susan Wigdal ,  Keith V. Wood ,  Monika G. Wood

    发明人： Brock Binkowski ,  Frank Fan ,  Susan Wigdal ,  Keith V. Wood ,  Monika G. Wood

    IPC分类号： C12Q1/68 ,  C07H21/00 ,  C12N15/63 ,  C12N5/10 ,  C12N9/02 ,  C12Q1/02 ,  C12Q1/66

    CPC分类号： C12N9/0069 ,  C12N9/0004 ,  C12Q1/66 ,  C12Y113/12007 ,  G01N33/84

    摘要： A modified luciferase protein which is a sensor for molecules including cAMP, cGMP, calcium, chelators thereof, kinases, or phosphatases is provided. Also provided is a circularly permuted anthozoan luciferase protein and a decapod crustacean luciferase protein, optionally containing one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest. Further provided is a modified anthozoan luciferase protein and a decapod crustacean luciferase protein containing an insertion of one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest.

    摘要翻译： 提供了一种修饰的荧光素酶蛋白，其是用于分子的传感器，包括cAMP，cGMP，钙，其螯合剂，激酶或磷酸酶。 还提供了循环置换的anthozoan萤光素酶蛋白质和十足配甲壳素荧光素酶蛋白质，任选地含有一个或多个异源氨基酸序列，其中至少一个直接或间接与感兴趣的分子相互作用。 进一步提供了一种修饰的anthozoan萤光素酶蛋白质和十足类甲壳类荧光素酶蛋白质，其含有一个或多个异源氨基酸序列的插入，其中至少一个与感兴趣的分子直接或间接相互作用。