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    Method for Generating Stable Cell Lines Expressing High Levels of a Protein of Interest
    1.
    发明申请
    Method for Generating Stable Cell Lines Expressing High Levels of a Protein of Interest 审中-公开
    产生表达高水平蛋白质的稳定细胞系的方法

    公开(公告)号：US20100184090A1

    公开(公告)日：2010-07-22

    申请号：US12666434

    申请日：2008-07-16

    申请人： Tara Kristen Crawford McFadd ,  Meijia Yang

    发明人： Tara Kristen Crawford McFadd ,  Meijia Yang

    IPC分类号： G01N33/53 ,  C12P21/06

    CPC分类号： C12N15/65

    摘要： This invention relates to industrial production of proteins. More specifically, the invention relates to a method for obtaining cells that stably express a protein of interest, even when cultivated in the absence of selective pressure. DHFR is used as a surrogate marker. The transfected cells are not selected based on resistance to a toxic compound, but based on fluorescence as measured by FACS using fluorescent MTX.

    摘要翻译： 本发明涉及蛋白质的工业化生产。 更具体地说，本发明涉及获得稳定表达目的蛋白质的细胞的方法，即使在没有选择压力的情况下培养。 DHFR用作替代标记。 转染的细胞不是基于对有毒化合物的抗性而是基于通过使用荧光MTX的FACS测量的荧光来选择的。

    HsReq*1 and hsReq*2proteins and use thereof to detect CDK2
    3.
    发明授权
    HsReq*1 and hsReq*2proteins and use thereof to detect CDK2 失效
    HsReq * 1和hsReq * 2蛋白及其用于检测CDK2

    公开(公告)号：US06521412B1

    公开(公告)日：2003-02-18

    申请号：US09338125

    申请日：1999-06-22

    申请人： Meijia Yang ,  Krishnan Nandabalan ,  Vincent Peter Schulz

    发明人： Meijia Yang ,  Krishnan Nandabalan ,  Vincent Peter Schulz

    IPC分类号： C12Q102

    CPC分类号： C07K14/4738 ,  A01K2217/05 ,  A01K2217/075 ,  A61K38/00 ,  A61K48/00 ,  C07K2319/00

    摘要： The present invention relates to complexes of the CDK2 protein with proteins identified as interacting with CDK2 by a modified yeast two hybrid assay system. The proteins identified to interact with CDK2 are cyclin H, cyclin I, ERH, and two gene products, hsReq*-1 and hsReq*-2, which are splice variants of the gene hsReq. Thus, the invention provides complexes of CDK2 and cyclin H, cyclin I, ERH, hsReq*-1, and hsReq*-2, and derivatives, fragments and analogs thereof. The invention also provides nucleic acids encoding the hsReq*-1 and hsReq*-2, and proteins and derivatives, fragments and analogs thereof. Methods of screening the complexes for efficacy in treating and/or preventing certain diseases and disorders, particularly cancer, atherosclerosis and neurodegenerative disease are also provided.

    摘要翻译： 本发明涉及CDK2蛋白与通过修饰的酵母双杂交测定系统鉴定为与CDK2相互作用的蛋白质的复合物。 鉴定为与CDK2相互作用的蛋白质是细胞周期蛋白H，细胞周期蛋白I，ERH和两个基因产物hsReq * -1和hsReq * -2，它们是hsReq基因的剪接变体。 因此，本发明提供了CDK2和细胞周期蛋白H，细胞周期蛋白I，ERH，hsReq * -1和hsReq * -2及其衍生物，片段和类似物的复合物。 本发明还提供编码hsReq * -1和hsReq * -2的核酸，以及蛋白质及其衍生物，片段和类似物。 还提供了筛选复合物治疗和/或预防某些疾病和病症，特别是癌症，动脉粥样硬化和神经变性疾病的功效的方法。

    Identification and comparison of protein—protein interactions that occur in populations and identification of inhibitors of these interactors
    4.
    发明授权
    Identification and comparison of protein—protein interactions that occur in populations and identification of inhibitors of these interactors 失效
    识别和比较发生在群体中的蛋白质 - 蛋白质相互作用以及鉴定这些相互作用体的抑制剂

    公开(公告)号：US06410239B1

    公开(公告)日：2002-06-25

    申请号：US09461125

    申请日：1999-12-14

    申请人： Krishnan Nandabalan ,  Jonathan Marc Rothberg ,  Meijia Yang ,  James Robert Knight ,  Theodore Samuel Kalbfleisch

    发明人： Krishnan Nandabalan ,  Jonathan Marc Rothberg ,  Meijia Yang ,  James Robert Knight ,  Theodore Samuel Kalbfleisch

    IPC分类号： C12Q168

    CPC分类号： C40B30/04 ,  B01J2219/00605 ,  B01J2219/0061 ,  B01J2219/00659 ,  B01J2219/00677 ,  B01J2219/00689 ,  B01J2219/00695 ,  B82Y30/00 ,  C12N15/1055 ,  C12N15/81 ,  G01N33/6845 ,  G06F19/22

    摘要： Methods are described for detecting protein-protein interactions, among two populations of proteins, each having a complexity of at least 100. Encoded proteins are fused either to the DNA-binding domain of a transcriptional activator or to the activation domain of a transcriptional activator. Two yeast strains, of the opposite mating type and carrying one type each of the fusion proteins are mated together. Productive interactions between the two halves due to protein-protein interactions lead to the reconstitution of the transcriptional activator, which in turn leads to the activation of a reporter gene containing a binding site for the DNA-binding domain. This analysis can be carried out for two or more populations of proteins. The differences in the genes encoding the proteins involved in the protein-protein interactions are characterized, thus leading to the identification of specific protein-protein interactions, and the genes encoding the interacting proteins, relevant to a particular tissue, stage or disease. Furthermore, inhibitors that interfere with these protein-protein interactions are identified by their ability to inactivate a reporter gene. The screening for such inhibitors can be in a multiplexed format where a set of inhibitors will be screened against a library of interactors. Further, information-processing methods and systems are described. These methods and systems pro vide for identification of the genes coding for detected interacting proteins, for assembling a unified database of protein-protein interaction data, and for processing this unified database to obtain protein interaction domain and protein pathway information.

    摘要翻译： 描述了在两个蛋白质群体中检测蛋白质 - 蛋白质相互作用的方法，每个蛋白质的复杂度至少为100.编码的蛋白质与转录激活物的DNA结合结构域或转录激活子的活化结构域融合。 具有相反交配型并携带一种类型的每种融合蛋白的两个酵母菌株配合在一起。 由于蛋白质 - 蛋白质相互作用，两半之间的生产性相互作用导致转录激活物的重组，其又导致含有DNA结合结构域的结合位点的报告基因的活化。 可以对两个或更多个蛋白质群体进行该分析。 表征编码参与蛋白质 - 蛋白质相互作用的蛋白质的基因的差异，从而导致鉴定特异性蛋白质 - 蛋白质相互作用，以及编码与特定组织，阶段或疾病相关的相互作用蛋白质的基因。 此外，干扰这些蛋白质 - 蛋白质相互作用的抑制剂通过其使报告基因失活的能力来鉴定。 这种抑制剂的筛选可以是复合形式，其中一组抑制剂将针对相互作用物质库进行筛选。 此外，描述信息处理方法和系统。 这些方法和系统提供了编码检测到的相互作用蛋白的基因的鉴定，用于组装蛋白质 - 蛋白质相互作用数据的统一数据库，并且用于处理该统一数据库以获得蛋白质相互作用域和蛋白质途径信息。

    53BP2 complexes
    5.
    发明授权
    53BP2 complexes 失效
    53BP2复合物

    公开(公告)号：US06627405B1

    公开(公告)日：2003-09-30

    申请号：US09338123

    申请日：1999-06-22

    申请人： Krishnan Nandabalan ,  Meijia Yang ,  Vincent Peter Schulz

    发明人： Krishnan Nandabalan ,  Meijia Yang ,  Vincent Peter Schulz

    IPC分类号： G01N3353

    CPC分类号： C07K14/47 ,  A01K2207/15 ,  A01K2217/00 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2267/03 ,  A61K38/00 ,  A61K48/00 ,  C07K2319/00 ,  C12N15/8509

    摘要： The present invention relates to complexes of the 53BP2 protein with proteins identified as interacting with 53BP2 by a yeast two hybrid assay system. The proteins identified to interact with 53BP2 are &bgr;-tubulin, p62, hnRNP G, and three gene products, 53BP2-IP1, 53BP2-IP2, and 53BP2-IP3 encoded, in part, by the EST R72810 sequence. Thus, the invention provides complexes of 53BP2 and &bgr;-tubulin, p62, hnRNP G, 53BP2-IP1, 53BP2-IP2, and 53BP2-IP3 and derivatives, fragments and analogs thereof. The invention also provides the 53BP2-IP1, 53BP2-IP2 and 53BP2-IP3 genes and proteins and derivatives, fragments and analogs thereof. Methods of screening the complexes for efficacy in treating and/or preventing certain diseases and disorders, particularly cancer, autoimmune disease and neurodegenerative disease are also provided.

    摘要翻译： 本发明涉及53BP2蛋白与通过酵母双杂交测定系统鉴定为与53BP2相互作用的蛋白质的复合物。 鉴定为与53BP2相互作用的蛋白质是β-微管蛋白，p62，hnRNP G和三种基因产物，53BP2-IP1,53BP2-IP2和53BP2-IP3，部分由EST R72810序列编码。 因此，本发明提供了53BP2和β-微管蛋白，p62，hnRNP G，53BP2-IP1,53BP2-IP2和53BP2-IP3的复合物及其衍生物，片段和类似物。 本发明还提供了53BP2-IP1,53BP2-IP2和53BP2-IP3基因及其蛋白质及其衍生物，片段和类似物。 还提供了筛选复合物治疗和/或预防某些疾病和病症，特别是癌症，自身免疫性疾病和神经变性疾病的功效的方法。

    Isolation and characterization of Hermansky Pudlak Syndrome (HPS) protein complexes and HPS protein-interacting proteins
    6.
    发明授权
    Isolation and characterization of Hermansky Pudlak Syndrome (HPS) protein complexes and HPS protein-interacting proteins 失效
    Hermansky Pudlak综合征（HPS）蛋白复合物和HPS蛋白相互作用蛋白的分离和表征

    公开(公告)号：US06573364B1

    公开(公告)日：2003-06-03

    申请号：US09266225

    申请日：1999-03-10

    申请人： Krishnan Nandabalan ,  Meijia Yang

    发明人： Krishnan Nandabalan ,  Meijia Yang

    IPC分类号： C07K1447

    CPC分类号： C12N9/1205 ,  A61K38/00 ,  C07K14/47 ,  C07K2319/00 ,  G01N33/6893 ,  G01N2800/207 ,  G01N2800/52

    摘要： The present invention relates to complexes of the HPS protein with proteins identified as interacting with HPS protein by a modified yeast two hybrid assay system. The proteins identified to interact with HPS protein are 14-3-3 protein, Hrs, atrophin-1, DGS-I, nuclear factor NF90, HPIP1 and human HN1 homolog protein. Accordingly, the present invention discloses complexes of HPS protein and 14-3-3 protein, Hrs, atrophin-1, DGS-I, nuclear factor NF90, HPIP1 and human HN1 homolog protein, and derivatives, fragments and analogs thereof. Additionally, the present invention also discloses nucleic acids encoding the HPIP1 and human HN1 homolog protein, or derivatives, fragments and analogs thereof. Methods of screening the complexes or proteins for efficacy in treating and/or preventing certain diseases and disorders, particularly atopic diseases, autoimmune diseases, neurodegenerative disease, cancer, pigmentation disorders, platelet dysfunction and viral diseases, are also disclosed herein.

    摘要翻译： 本发明涉及HPS蛋白与通过修饰的酵母双杂交测定系统鉴定为与HPS蛋白相互作用的蛋白质的复合物。 鉴定与HPS蛋白相互作用的蛋白质是14-3-3蛋白，Hrs，atrophin-1，DGS-1，核因子NF90，HPIP1和人HN1同源蛋白。 因此，本发明公开了HPS蛋白和14-3-3蛋白，Hrs，atrophin-1，DGS-1，核因子NF90，HPIP1和人HN1同源蛋白的复合物及其衍生物，片段和类似物。 此外，本发明还公开了编码HPIP1和人HN1同源蛋白或其衍生物，片段和类似物的核酸。 本文还公开了筛选复合物或蛋白质以治疗和/或预防某些疾病和障碍，特别是特应性疾病，自身免疫疾病，神经变性疾病，癌症，色素沉着病症，血小板功能障碍和病毒性疾病的功效的方法。

    Identification and comparison of protein-protein interactions that occur in populations and identification of inhibitors of these interactors
    7.
    发明授权
    Identification and comparison of protein-protein interactions that occur in populations and identification of inhibitors of these interactors 失效
    识别和比较发生在群体中的蛋白质 - 蛋白质相互作用以及鉴定这些相互作用体的抑制剂

    公开(公告)号：US6057101A

    公开(公告)日：2000-05-02

    申请号：US874825

    申请日：1997-06-13

    申请人： Krishnan Nandabalan ,  Jonathan Marc Rothberg ,  Meijia Yang ,  James Robert Knight ,  Theodore Samuel Kalbfleisch

    发明人： Krishnan Nandabalan ,  Jonathan Marc Rothberg ,  Meijia Yang ,  James Robert Knight ,  Theodore Samuel Kalbfleisch

    IPC分类号： C12N1/15 ,  C12N15/10 ,  C12N15/81 ,  C12P21/06 ,  C12Q1/68 ,  C40B30/04 ,  G01N33/68 ,  G06F17/30 ,  C12Q1/70

    CPC分类号： C40B30/04 ,  B82Y30/00 ,  C12N15/1055 ,  C12N15/81 ,  G01N33/6845 ,  B01J2219/00605 ,  B01J2219/0061 ,  B01J2219/00659 ,  B01J2219/00677 ,  B01J2219/00689 ,  B01J2219/00695 ,  G06F19/22

    摘要： Methods are described for detecting protein-protein interactions, among two populations of proteins, each having a complexity of at least 1,000. For example, proteins are fused either to the DNA-binding domain of a transcriptional activator or to the activation domain of a transcriptional activator. Two yeast strains, of the opposite mating type and carrying one type each of the fusion proteins are mated together. Productive interactions between the two halves due to protein-protein interactions lead to the reconstitution of the transcriptional activator, which in turn leads to the activation of a reporter gene containing a binding site for the DNA-binding domain. This analysis can be carried out for two or more populations of proteins. The differences in the genes encoding the proteins involved in the protein-protein interactions are characterized, thus leading to the identification of specific protein-protein interactions, and the genes encoding the interacting proteins, relevant to a particular tissue, stage or disease. Furthermore, inhibitors that interfere with these protein-protein interactions are identified by their ability to inactivate a reporter gene. The screening for such inhibitors can be in a multiplexed format where a set of inhibitors will be screened against a library of interactors. Further, information-processing methods and systems are described. These methods and systems provide for identification of the genes coding for detected interacting proteins, for assembling a unified database of protein-protein interaction data, and for processing this unified database to obtain protein interaction domain and protein pathway information.

    摘要翻译： 描述了在两个蛋白质群体中检测蛋白质 - 蛋白质相互作用的方法，每个蛋白质的复杂度至少为1,000。 例如，蛋白质融合到转录激活物的DNA结合域或转录激活子的激活结构域。 具有相反交配型并携带一种类型的每种融合蛋白的两个酵母菌株配合在一起。 由于蛋白质 - 蛋白质相互作用，两半之间的生产性相互作用导致转录激活物的重组，其又导致含有DNA结合结构域的结合位点的报告基因的活化。 可以对两个或更多个蛋白质群体进行该分析。 表征编码参与蛋白质 - 蛋白质相互作用的蛋白质的基因的差异，从而导致鉴定特异性蛋白质 - 蛋白质相互作用，以及编码与特定组织，阶段或疾病相关的相互作用蛋白质的基因。 此外，干扰这些蛋白质 - 蛋白质相互作用的抑制剂通过其使报告基因失活的能力来鉴定。 这种抑制剂的筛选可以是复合形式，其中一组抑制剂将针对相互作用物质库进行筛选。 此外，描述信息处理方法和系统。 这些方法和系统提供了编码检测到的相互作用蛋白的基因的鉴定，用于组装蛋白质 - 蛋白质相互作用数据的统一数据库，并且用于处理该统一数据库以获得蛋白质相互作用区域和蛋白质途径信息。