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    Binary signal detection assays
    41.
    发明申请
    Binary signal detection assays 审中-公开
    二进制信号检测测定

    公开(公告)号：US20080050743A1

    公开(公告)日：2008-02-28

    申请号：US11821286

    申请日：2007-06-22

    Applicant: Joseph Sorge ,  Carsten-Peter Carstens ,  Craig Monell ,  Alexander Belyaev

    Inventor： Joseph Sorge ,  Carsten-Peter Carstens ,  Craig Monell ,  Alexander Belyaev

    IPC: C12Q1/68

    CPC classification number: C12Q1/6816 ,  C12Q2565/201 ,  C12Q2521/301 ,  C12Q2521/337

    Abstract: The present invention provides methods, kits and compositions for the detection of an analyte. The invention is particularly suited for the detection and quantification of analytes in solution. In the methods of the invention a complex is formed between two or more analyte specific probes (ASP) and an analyte. The reactive moieties of the probes interact upon the binding of the analyte specific probes to the analyte. The reactive moieties generate a nucleic acid cleavage product which is detected and indicative of the presence of the analyte.

    Abstract translation: 本发明提供了用于检测分析物的方法，试剂盒和组合物。 本发明特别适用于溶液中分析物的检测和定量。 在本发明的方法中，在两种或多种分析物特异性探针（ASP）和分析物之间形成复合物。 探针的反应性部分在分析物特异性探针与分析物结合时相互作用。 反应性部分产生核酸切割产物，其被检测并指示分析物的存在。

    Methods and compositions for interaction trap assays
    43.
    发明授权
    Methods and compositions for interaction trap assays 有权
    相互作用阱测定法的方法和组成

    公开(公告)号：US07029847B2

    公开(公告)日：2006-04-18

    申请号：US09990762

    申请日：2001-11-14

    Applicant: J. Keith Joung ,  Jeffrey Miller ,  Carl O. Pabo

    Inventor： J. Keith Joung ,  Jeffrey Miller ,  Carl O. Pabo

    IPC: C12Q1/68 ,  G01N33/53 ,  G01N33/567

    CPC classification number: C12N15/1055 ,  C12Q1/6897 ,  G01N33/50 ,  G01N33/5008 ,  G01N33/502 ,  C12Q2565/201

    Abstract: The present invention provides methods and compositions for interaction trap assays for detecting protein-protein, protein-DNA, or protein-RNA interactions. The methods and compositions of the invention may also be used to identify agents which may agonize or antagonize a protein-protein, protein-DNA, or protein-RNA interaction. In certain embodiments, the interaction trap system of the invention is useful for screening libraries with greater than 107 members. In other embodiments, the interaction trap system of the invention is used in conjunction with flow cytometry. The invention further provides a means for simultaneously screening a target protein or nucleic acid sequence for the ability to interact with two or more test proteins or nucleic acids.

    Abstract translation: 本发明提供了用于检测蛋白质 - 蛋白质，蛋白质-DNA或蛋白质 - RNA相互作用的相互作用阱测定法的方法和组合物。 本发明的方法和组合物还可用于鉴定可能激动或拮抗蛋白质 - 蛋白质，蛋白质-DNA或蛋白质 - RNA相互作用的试剂。 在某些实施方案中，本发明的相互作用捕获系统可用于筛选具有大于10个成员的文库。 在其它实施方案中，本发明的相互作用阱系统结合流式细胞术使用。 本发明还提供了用于同时筛选靶蛋白或核酸序列以获得与两种或多种测试蛋白或核酸相互作用的能力的方法。

    Genetic assay for protein nuclear transport
    44.
    发明授权
    Genetic assay for protein nuclear transport 失效
    蛋白质核转运的遗传测定

    公开(公告)号：US06902886B1

    公开(公告)日：2005-06-07

    申请号：US09435274

    申请日：1999-11-05

    Applicant: Vitaly H. Citovsky ,  Yoon Rhee

    Inventor： Vitaly H. Citovsky ,  Yoon Rhee

    IPC: C07H21/04 ,  C07K14/195 ,  C12Q1/68

    CPC classification number: C07K14/195 ,  C07H21/04 ,  C12Q1/6897 ,  C12Q2565/201

    Abstract: The invention provides methods of determining the presence of a nuclear localization signal and/or the presence of a nuclear export signal in a protein of interest. The invention further provides chimeric nucleic acids and recombinant host cells for use in such methods. Additionally provided is a nucleic acid molecule encoding a modified LexA protein, wherein the modified LexA protein has no nuclear localization signal, as well as the modified LexA protein itself. In the nuclear import assay, if a protein of interest fused to a mLexA-Gal4AD hybrid contains a functional NLS, the fusion product will enter the yeast cell nucleus and activate the expression of reporter genes. In the nuclear export assay, if a protein of interest fused to a mLexA-SV40 NLS-Gal4AD hybrid contains a functional NES, the fusion product localized to the cell nucleus will exit into the cytoplasm, decreasing the reporter gene expression levels.

    Abstract translation: 本发明提供确定核定位信号的存在和/或感兴趣的蛋白质中核出口信号的存在的方法。 本发明还提供了用于这些方法的嵌合核酸和重组宿主细胞。 另外提供了编码修饰的LexA蛋白质的核酸分子，其中修饰的LexA蛋白质没有核定位信号，以及修饰的LexA蛋白质本身。 在核进口测定中，如果与mLexA-Gal 4 AD杂交融合的感兴趣的蛋白质含有功能性NLS，则融合产物将进入酵母细胞核并激活报道基因的表达。 在核出口测定中，如果与mLexA-SV 40 NLS-Gal 4 AD杂交融合的感兴趣的蛋白质含有功能性NES，则定位于细胞核的融合产物将退出细胞质，降低报道基因表达水平。

    Mammalian two-hybrid system for screening for modulators of the accumulation of metabolic products
    46.
    发明申请
    Mammalian two-hybrid system for screening for modulators of the accumulation of metabolic products 有权
    哺乳动物双杂交系统用于筛选代谢产物积累的调节剂

    公开(公告)号：US20040265791A1

    公开(公告)日：2004-12-30

    申请号：US10896600

    申请日：2004-07-21

    Applicant: The Regents of the University of California

    Inventor： Osamu Tetsu ,  Kenichi Wakita ,  Frank McCormick

    IPC: C12Q001/00 ,  C12Q001/68

    CPC classification number: C12N15/1055 ,  A61K48/00 ,  A61K48/0058 ,  C12Q1/6897 ,  C12Q2565/201

    Abstract: This invention utilizes a two-hybrid system to screen for agents that modulate the ability of a cell to degrade or to accumulate a metabolic product or to selective kill a cell or to selectively express a gene or cDNA in a cell that has a defect in its ability to degrade or to accumulate a metabolic product. One embodiment provides a mammalian cell comprising a nucleic acid encoding a peptide binding domain and an effector gene; a first chimeric protein comprising a nucleic acid binding domain that binds the peptide binding domain attached to the metabolic product or to a ligand that binds to the metabolic product; and a second chimeric protein comprising an expression control protein attached to the metabolic product or to the ligand that binds to the metabolic product such that when the first chimeric protein comprises the metabolic product, the second chimeric protein comprises the ligand and when the first chimeric protein comprises the ligand, the second chimeric protein comprises the metabolic product. The cell is contacted with a test agent and an alteration of expression of the effector gene is detected (if present) where a change in expression of the effector gene in response to the test agent indicates that the test agent modulates the ability of the cell to accumulate or degrade the metabolic product.

    Assembly and screening of highly complex and fully human antibody repertoire in yeast
    49.
    发明授权
    Assembly and screening of highly complex and fully human antibody repertoire in yeast 有权
    组装和筛选酵母中高度复杂和完全的人体抗体谱

    公开(公告)号：US06610472B1

    公开(公告)日：2003-08-26

    申请号：US09703399

    申请日：2000-10-31

    Applicant: Li Zhu ,  Shaobing Benjamin Hua ,  James Sheridan ,  Yu-Huei Lin

    Inventor： Li Zhu ,  Shaobing Benjamin Hua ,  James Sheridan ,  Yu-Huei Lin

    IPC: G01N3353

    CPC classification number: C40B30/04 ,  C07K16/00 ,  C07K2317/21 ,  C07K2317/55 ,  C12Q1/6897 ,  G01N33/6845 ,  C12Q2565/201

    Abstract: Compositions, methods, and kits are provided for efficiently generating and screening a library of highly diverse protein complexes for their ability to bind to other proteins or oligonucleotide sequences. In one aspect of the invention, a library of expression vectors is provided for expressing the library of protein complexes. The library comprises a first nucleotide sequence encoding a first polypeptide subunit; and a second nucleotide sequence encoding a second polypeptide subunit. The first and second nucleotide sequences each independently varies within the library of expression vectors. In addition, the first and second polypeptide subunit are expressed as separate proteins which self-assemble to form a protein complex, such as a double-chain antibody fragment (dcFv or Fab) and a fully assembled antibody, in cells into which the library of expression vectors are introduced. The library of expression vectors can be efficiently generated in yeast cells through homologous recombination; and the encoded proteins complexes with high binding affinity to their target molecule can be selected by high throughput screening in vivo or in vitro.

    Abstract translation: 提供了组合物，方法和试剂盒，用于有效地产生和筛选高度多样化的蛋白质复合物的文库以获得与其它蛋白质或寡核苷酸序列结合的能力。 在本发明的一个方面，提供表达载体文库用于表达蛋白质复合物的文库。 该文库包含编码第一多肽亚基的第一核苷酸序列; 和编码第二多肽亚基的第二核苷酸序列。 第一和第二核苷酸序列各自独立地在表达载体的文库内变化。 此外，第一和第二多肽亚基表达为分离的蛋白，其自组装形成蛋白复合物，例如双链抗体片段（dcFv或Fab）和完全组装的抗体，其中文库 引入表达载体。 表达载体文库可以通过同源重组在酵母细胞中有效产生; 并且可以通过体内或体外的高通量筛选来选择对其靶分子具有高结合亲和力的编码蛋白复合物。

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