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    Fluorescent proteins
    1.
    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。

    Method for extracting quantitative information relating to interactions between cellular components
    2.
    发明授权
    Method for extracting quantitative information relating to interactions between cellular components 有权
    提取与细胞成分之间相互作用有关的定量信息的方法

    公开(公告)号：US07282347B2

    公开(公告)日：2007-10-16

    申请号：US10332065

    申请日：2001-07-03

    申请人： Sara Petersen Bjorn ,  Ole Thastrup ,  Bernard Robert Terry ,  Grith Hagel ,  Soren Jensby Nielsen

    发明人： Sara Petersen Bjorn ,  Ole Thastrup ,  Bernard Robert Terry ,  Grith Hagel ,  Soren Jensby Nielsen

    IPC分类号： C07H21/04 ,  C12Q1/68 ,  C12N9/12 ,  C12N5/06

    CPC分类号： G01N33/6845 ,  G01N33/5008 ,  G01N33/502 ,  G01N33/5032 ,  G01N33/68

    摘要： A method is described to assay for protein interactions in living cells, e.g. by the introduction of two heterologous conjugates into the cell. The method uses the measurement of cellular distribution of a detectable component (e.g. a GFP-labeled˜fluorescent probe) to indicate the presence or absence of an interaction between that component and a second component of interest. The method uses the knowledge that certain components can be stimulated to redistribute within the cell to defined locations. Inducible redistribution systems make it possible to determine if specific interactions occur between components. Inducible systems are described where it is demonstrated that the redistribution stimuli are essentially “null”, in that they affect no other system in the cell during the assay period, other than the component whose redistribution can be induced. Also described is an extraction buffer which is useful in high throughput screening for drugs which affect the intracellular distribution of intracellular components. The extraction buffer comprises a cellular fixation agent and cellular permeabilisation agent. Optimizing the composition of the extraction buffer and its application to various cell types is described.

    摘要翻译： 描述了一种测定活细胞中蛋白质相互作用的方法，例如 通过将两种异源偶联物引入细胞。 该方法使用可检测组分（例如GFP-标记的〜荧光探针）的细胞分布的测量来指示该组分与感兴趣的第二组分之间存在或不存在相互作用。 该方法使用某些组件可以被激励以在单元格内重新分配到定义的位置的知识。 诱导再分配系统可以确定组件之间是否发生特定的相互作用。 描述了可诱导的系统，其中证明再分布刺激基本上是“无效”的，因为除了可以诱导其再分配的组分之外，它们在测定期间不影响细胞中的其它系统。 还描述了一种提取缓冲液，其可用于影响细胞内组分细胞内分布的药物的高通量筛选。 提取缓冲液包含细胞固定剂和细胞透化剂。 描述了优化提取缓冲液的组成及其在各种细胞类型中的应用。

    METHOD FOR EXTRACTING QUANTITATIVE INFORMATION RELATING TO AN INFLUENCE ON A CELLULAR RESPONSE
    3.
    发明申请
    METHOD FOR EXTRACTING QUANTITATIVE INFORMATION RELATING TO AN INFLUENCE ON A CELLULAR RESPONSE 审中-公开
    提取与细胞响应有关的量化信息的方法

    公开(公告)号：US20090023598A1

    公开(公告)日：2009-01-22

    申请号：US11927246

    申请日：2007-10-29

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

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

    IPC分类号： C40B30/06

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

    摘要： A method for screening a library of compounds to detect a biologically active compound that modulates intracellular translocation of a subunit of a component of an intracellular pathway affecting intracellular processes includes: culturing one or more cells containing a nucleotide sequence coding for a hybrid polypeptide comprising a luminophore linked to the subunit of the component; introducing a compound of the library of compounds into the cell culture; screening the compound to determine whether the compound modulates the intracellular translocation of the subunit of the component; measuring light emitted from the luminophore to determine a first distribution; measuring light emitted from the luminophore to determine a second distribution; computing a variation between the first distribution and the second distribution by processing the measured light, any variation is indicative that the compound is biologically active. The method is also performed with a library of compounds.

    摘要翻译： 筛选化合物文库以检测调节影响细胞内过程的细胞内途径的组分的亚单位的细胞内易位的生物活性化合物的方法包括：培养含有编码包含发光体的杂合多肽的核苷酸序列的一个或多个细胞 连接到组件的子单元; 将化合物的化合物引入细胞培养物中; 筛选化合物以确定化合物是否调节组分亚单位的细胞内易位; 测量从发光体发射的光以确定第一分布; 测量从发光体发射的光以确定第二分布; 通过处理测量的光来计算第一分布和第二分布之间的变化，任何变化表明化合物具有生物活性。 该方法也用化合物文库进行。

    Nucleic acids encoding fluorescent proteins and methods of using the same
    4.
    发明授权
    Nucleic acids encoding fluorescent proteins and methods of using the same 有权
    编码荧光蛋白的核酸及其使用方法

    公开(公告)号：US07476518B2

    公开(公告)日：2009-01-13

    申请号：US11206904

    申请日：2005-08-19

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

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

    IPC分类号： C12P21/06

    CPC分类号： C07K14/43595 ,  C07K2319/00

    摘要： A GFP with an F64L mutation and 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 clearer band separation when used together with those GFPs. Examples include the sequences in SEQ ID NOs: 3 and 4.

    摘要翻译： 提供了具有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。 实例包括SEQ ID NO：3和4中的序列。

    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。