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    Method of detecting biologically active substances
    3.
    发明授权
    Method of detecting biologically active substances 有权
    检测生物活性物质的方法

    公开(公告)号：US06566083B1

    公开(公告)日：2003-05-20

    申请号：US09346946

    申请日：1999-07-02

    申请人： Ole Thastrup ,  Søren Tullin ,  Lars Kongsbak Poulsen ,  Sara Petersen Bjørn

    发明人： Ole Thastrup ,  Søren Tullin ,  Lars Kongsbak Poulsen ,  Sara Petersen Bjørn

    IPC分类号： G01N33573

    CPC分类号： C07K14/43595 ,  C07K2319/00 ,  C12N15/65 ,  G01N33/5005 ,  G01N33/533 ,  G01N33/542 ,  G01N2333/43595 ,  G01N2333/9121

    摘要： The present invention relates to a method of detecting biologically active substances affecting intracellular processes, an isolated DNA, a vector and a cell for use in the method. More specifically, the present invention provides a method for detecting a biologically active substance that affects intracellular processes mediated through a protein kinase or a second messenger, which comprises incubating a cell with a test substance and measuring any change caused by the test substance in the flourescence of (i) a wild-type or modified green flourescent protein (GFP) having a protein kinase recognition site, (ii) a modified GFP containing a second messenger binding domain, or (iii) a hybrid polypeptide having a wild-type or modified GFP and an attached protein kinase recognition site or a second messenger binding domain.

    摘要翻译： 本发明涉及检测影响细胞内过程的生物活性物质的方法，分离的DNA，载体和用于该方法的细胞。 更具体地，本发明提供了一种检测影响通过蛋白激酶或第二信使介导的细胞内过程的生物活性物质的方法，其包括将细胞与测试物质一起孵育并测量由荧光检测物质引起的任何变化 （i）具有蛋白激酶识别位点的野生型或修饰型绿色荧光蛋白（GFP），（ii）含有第二信使结合结构域的修饰型GFP，或（iii）具有野生型或修饰型的杂合多肽 GFP和附着的蛋白激酶识别位点或第二信使结合结构域。

    Compounds modifying apoptosis
    7.
    发明授权
    Compounds modifying apoptosis 有权
    化合物修饰凋亡

    公开(公告)号：US08318717B2

    公开(公告)日：2012-11-27

    申请号：US11914994

    申请日：2006-05-23

    申请人： Ole Thastrup ,  Jens Chr. Norrild

    发明人： Ole Thastrup ,  Jens Chr. Norrild

    IPC分类号： A61K31/397 ,  C07C233/00 ,  C07D207/16

    CPC分类号： C07C237/22 ,  C07B2200/11 ,  C07D207/16 ,  C07D211/64

    摘要： The present invention relates to compounds capable of inhibiting binding of the Smac protein to Inhibitors of apoptosis (IAPs). Such compounds are preferably capable of inhibiting IAP and thus may promote apoptosis or sensitize cells for apoptosis. The compounds may be used in the treatment of proliferative diseases, such as cancer.

    摘要翻译： 本发明涉及能够抑制Smac蛋白与凋亡抑制剂（IAP）结合的化合物。 这些化合物优选能够抑制IAP，从而可以促进细胞凋亡或使细胞增殖以进行细胞凋亡。 该化合物可用于治疗增殖性疾病，例如癌症。

    Method for extracting quantitative information relating to an influence on a cellular response
    8.
    发明授权
    Method for extracting quantitative information relating to an influence on a cellular response 有权
    提取与细胞反应有关的定量信息的方法

    公开(公告)号：US08058008B2

    公开(公告)日：2011-11-15

    申请号：US10072036

    申请日：2002-02-05

    申请人： Ole Thastrup ,  Sara Petersen Bjørn ,  Soren Tullin ,  Kasper Almholt ,  Kurt Scudder

    发明人： Ole Thastrup ,  Sara Petersen Bjørn ,  Soren Tullin ,  Kasper Almholt ,  Kurt Scudder

    IPC分类号： G01N33/53 ,  G01N21/76 ,  C12P21/02

    CPC分类号： G01N33/5041 ,  G01N33/5005 ,  G01N33/5008 ,  G01N33/5014 ,  G01N33/5035 ,  G01N33/533 ,  G01N33/582 ,  G01N2333/43595 ,  G01N2333/9121

    摘要： Cells are genetically modified to express a luminophore, e.g., a modified (F64L, S65T, Y66H) Green Fluorescent Protein (GFP, EGFP) coupled to a component of an intracellular signalling pathway such as a transcription factor, a cGMP- or cAMP-dependent protein kinase, a cyclin-, calmodulin- or phospholipid-dependent or mitogen-activated serine/threonin protein kinase, a tyrosine protein kinase, or a protein phosphatase (e.g. PKA, PKC, Erk, Smad, VASP, actin, p38, Jnk1, PKG, IkappaB, CDK2, Grk5, Zap70, p85, protein-tyrosine phosphatase 1C, Stat5, NFAT, NFkappaB, RhoA, PKB). An influence modulates the intracellular signalling pathway in such a way that the luminophore is being redistributed or translocated with the component in living cells in a manner experimentally determined to be correlated to the degree of the influence. Measurement of redistribution is performed by recording of light intensity, fluorescence lifetime, polarization, wavelength shift, resonance energy transfer, or other properties by an apparatus consisting of e.g. a fluorescence microscope and a CCD camera. Data stored as digital images are processed to numbers representing the degree of redistribution. The method can be used as a screening program for identifying a compound that modulates a component and is capable of treating a disease related to the function of the component.

    摘要翻译： 细胞被遗传修饰以表达发光体，例如，与细胞内信号传导途径的组分偶联的经修饰的（F64L，S65T，Y66H）绿色荧光蛋白（GFP，EGFP），例如转录因子，cGMP-或cAMP依赖性 蛋白激酶，细胞周期蛋白，钙调蛋白或磷脂依赖性或丝裂原活化的丝氨酸/苏氨酸蛋白激酶，酪氨酸蛋白激酶或蛋白磷酸酶（例如PKA，PKC，Erk，Smad，VASP，肌动蛋白，p38，Jnk1， PKG，IkappaB，CDK2，Grk5，Zap70，p85，蛋白质 - 酪氨酸磷酸酶1C，Stat5，NFAT，NFkappaB，RhoA，PKB）。 影响调节细胞内信号传导途径，使得发光体在活体细胞中以组分重新分配或移位，其方式通过实验确定与影响程度相关。 重新分布的测量是通过记录光强度，荧光寿命，极化，波长偏移，共振能量转移或其它性质通过由例如， 荧光显微镜和CCD相机。 作为数字图像存储的数据被处理成表示再分配程度的数字。 该方法可以用作鉴定调节成分并能够治疗与该成分的功能有关的疾病的化合物的筛选程序。

    Fluorescent proteins
    9.
    发明授权
    Fluorescent proteins 有权
    荧光蛋白

    公开(公告)号：US07001986B2

    公开(公告)日：2006-02-21

    申请号：US09887784

    申请日：2001-06-19

    申请人： Sara Petersen Bjorn ,  Len Pagliaro ,  Ole Thastrup

    发明人： Sara Petersen Bjorn ,  Len Pagliaro ,  Ole Thastrup

    IPC分类号： C07K1/00

    CPC分类号： C07K14/43595 ,  C07K2319/00

    摘要： A GFP with an F64L mutation and an E222G mutation is provided. This GFP has a bigger Stokes shift compared to other GFPs making it very suitable for high throughput screening due to a better resolution. This GFP also has an excitation maximum between the yellow GFP and the cyan GFP allowing for cleaner band separation when used together with those GFPs.

    摘要翻译： 提供具有F64L突变和E222G突变的GFP。 与其他GFP相比，该GFP具有更大的斯托克斯转变，因此由于更好的分辨率，它非常适合高通量筛选。 该GFP还具有黄色GFP和青色GFP之间的激发最大值，当与那些GFP一起使用时，可以实现更清洁的带分离。