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。

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。