公开(公告)号：WO1996002561A1

公开(公告)日：1996-02-01

申请号：PCT/US1995009307

申请日：1995-07-20

Applicant: THE GENERAL HOSPITAL CORPORATION ,  GENETICS INSTITUTE, INC.

Inventor： THE GENERAL HOSPITAL CORPORATION ,  GENETICS INSTITUTE, INC. ,  BRENT, Roger ,  MCCOY, John, M. ,  JESSEN, Timm, H. ,  XU, Chanxing, Wilson

IPC: C07H21/04

CPC classification number: C12N9/0036 ,  C07K2319/00 ,  C07K2319/35 ,  C07K2319/70 ,  C07K2319/71 ,  C07K2319/80 ,  C12N15/1055 ,  C12Q1/6897

Abstract: Disclosed herein is a method of determining whether a first protein is capable of physically interacting with a second protein, involving: (a) providing a host cell which contains (i) a reporter gene operably linked to a protein binding site; (ii) a first fusion gene which expresses a first fusion protein, the first fusion protein including the first protein covalently bonded to a binding moiety which is capable of specifically binding to the protein binding site; and (iii) a second fusion gene which expresses a second fusion protein, the second fusion protein including the second protein covalently bonded to a gene activating moiety and being conformationally-constrained; and (b) measuring expression of the reporter gene as a measure of an interaction between the first and the second proteins. Also disclosed are methods for assaying protein interactions, and identifying antagonists and agonists of protein interactions. Proteins isolated by these methods are also discussed. Finally, populations of eukaryotic cells are disclosed, each cell having a recombinant DNA molecule encoding a conformationally-constrained intracellular peptide.

Abstract translation: 本文公开了一种确定第一蛋白质是否能够与第二蛋白质物理相互作用的方法，涉及：（a）提供宿主细胞，其包含（i）可操作地连接到蛋白质结合位点的报告基因; （ii）表达第一融合蛋白的第一融合基因，所述第一融合蛋白包括共价键合到能够特异性结合蛋白结合位点的结合部分的第一蛋白; 和（iii）表达第二融合蛋白的第二融合基因，所述第二融合蛋白包括共价键合到基因激活部分并被构象约束的第二蛋白; 和（b）测量报告基因的表达作为第一和第二蛋白质之间相互作用的量度。 还公开了用于测定蛋白质相互作用的方法，以及鉴定蛋白质相互作用的拮抗剂和激动剂。 还讨论了通过这些方法分离的蛋白质。 最后，公开了真核细胞群，每个细胞具有编码构象约束细胞内肽的重组DNA分子。

公开(公告)号：WO1994010300A1

公开(公告)日：1994-05-11

申请号：PCT/US1993010069

申请日：1993-10-20

Applicant: THE GENERAL HOSPITAL CORPORATION

Inventor： THE GENERAL HOSPITAL CORPORATION ,  BRENT, Roger ,  GYURIS, Jeno ,  GOLEMIS, Erica

IPC: C12N15/10

CPC classification number: C12Q1/6897 ,  A61K38/00 ,  C07K14/4738 ,  C07K2319/00 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N9/1205 ,  C12N9/16 ,  C12N15/1055 ,  C12Q2565/201

Abstract: Disclosed is a method for determining whether a first protein is capable of physically interacting with a second protein. The method involves: (a) providing a host cell which contains (i) a reporter gene operably linked to a protein binding site; (ii) a first fusion gene which expresses a first fusion protein, the first fusion protein including the first protein covalently bonded to a binding moiety which is capable of specifically binding to the protein binding site; and (iii) a second fusion gene which expresses a second fusion protein, the second fusion protein including the second protein covalently bonded to a weak gene activating moiety; and (b) measuring expression of the reporter gene as a measure of an interaction between the first and the second proteins. Such a determination facilitates the isolation of the gene encoding the interacting protein. Also disclosed herein is recombinant Cdi1 polypeptide, nucleic acid encoding the Cdi1 polypeptide, and uses thereof.

Abstract translation: 公开了一种用于确定第一蛋白质是否能够与第二蛋白质物理相互作用的方法。 该方法包括：（a）提供宿主细胞，其包含（i）与蛋白质结合位点可操作地连接的报告基因; （ii）表达第一融合蛋白的第一融合基因，所述第一融合蛋白包括共价键合到能够特异性结合蛋白结合位点的结合部分的第一蛋白; 和（iii）表达第二融合蛋白的第二融合基因，所述第二融合蛋白包括与弱基因活化部分共价结合的第二蛋白质; 和（b）测量报告基因的表达作为第一和第二蛋白质之间相互作用的量度。 这样的测定有助于分离编码相互作用蛋白的基因。 本文还公开了重组Cdi1多肽，编码Cdi1多肽的核酸及其用途。

公开(公告)号：WO1994017101A1

公开(公告)日：1994-08-04

申请号：PCT/US1993012643

申请日：1993-12-29

Applicant: THE GENERAL HOSPITAL CORPORATION

Inventor： THE GENERAL HOSPITAL CORPORATION ,  BRENT, Roger ,  ZERVOS, Antonis, S.

IPC: C07K13/00

CPC classification number: C07K14/47 ,  A61K38/00 ,  C12Q1/6886 ,  C12Q2600/136

Abstract: Disclosed are substantially pure preparations of Max-Interacting (Mxi) polypeptides, DNA encoding such polypeptides, antibodies recognizing such polypeptides, and diagnostic and therapeutic methods utilizing such polypeptides.

Abstract translation: 公开了最大相互作用（Mxi）多肽，编码这种多肽的DNA，识别这种多肽的抗体，以及利用这种多肽的诊断和治疗方法的基本上纯的制剂。

公开(公告)号：WO1997038127A1

公开(公告)日：1997-10-16

申请号：PCT/US1997005793

申请日：1997-04-09

Applicant: THE GENERAL HOSPITAL CORPORATION ,  GENETICS INSTITUTE, INC.

Inventor： THE GENERAL HOSPITAL CORPORATION ,  GENETICS INSTITUTE, INC. ,  BRENT, Roger ,  McCOY, John, M. ,  JESSEN, Timm, H. ,  XU, Chanxing Wilson

IPC: C12Q01/00

CPC classification number: C12Q1/025 ,  C07K7/08 ,  C07K14/00 ,  C07K2319/00 ,  C12N9/0036 ,  C12N15/1055 ,  C12Q1/6897 ,  C40B30/04 ,  G01N33/68 ,  G01N33/6845

Abstract: Disclosed herein is a method of determining whether a first protein is capable of physically interacting with a second protein, involving: (a) providing a host cell which contains (i) a reporter gene operably linked to a protein binding site; (ii) a first fusion gene which expresses a first fusion protein, the first fusion protein including the first protein covalently bonded to a binding moiety which is capable of specifically binding to the protein binding site; and (iii) a second fusion gene which expresses a second fusion protein, the second fusion protein including the second protein covalently bonded to a gene activating moiety and being conformationally-constrained; and (b) measuring expression of the reporter gene as a measure of an interaction between the first and the second proteins. Also disclosed are methods for assaying protein interactions, and identifying antagonists and agonists of protein interactions. Proteins isolated by these methods are also discussed. Finally, populations of eukaryotic cells are disclosed, each cell having a recombinant DNA molecule encoding a conformationally-constrained intracellular peptide.

Abstract translation: 本文公开了一种确定第一蛋白质是否能够与第二蛋白质物理相互作用的方法，涉及：（a）提供宿主细胞，其包含（i）可操作地连接到蛋白质结合位点的报告基因; （ii）表达第一融合蛋白的第一融合基因，所述第一融合蛋白包括共价键合到能够特异性结合蛋白结合位点的结合部分的第一蛋白; 和（iii）表达第二融合蛋白的第二融合基因，所述第二融合蛋白包括共价键合到基因激活部分并被构象约束的第二蛋白; 和（b）测量报告基因的表达作为第一和第二蛋白质之间相互作用的量度。 还公开了用于测定蛋白质相互作用的方法，以及鉴定蛋白质相互作用的拮抗剂和激动剂。 还讨论了通过这些方法分离的蛋白质。 最后，公开了真核细胞群，每个细胞具有编码构象约束细胞内肽的重组DNA分子。