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CO-TRANSLATIONAL ACTIVATION OF A TRANSCRIPTION FACTOR BY PORTEOLYTIC CLEAVAGE AND METHODS OF USE

机译:渗透裂解法对转录因子的共翻译活化及其使用方法

摘要

A method for measuring expression of autoregulatory molecules within living cells is provided. An autoregulatory molecule and marker construct is expressed in vivo, where the marker is cleaved from the construct during translation. The method comprises the expression of a construct having an autoregulatory molecule bound to a measurable expression marker by a cleavable linker. The cleavable linker is the substrate of a protease, which acts on its substrate in vivo during translation. Cleavage during translation, allows the autoregulatory molecule to fold normally as it would in its native form. The measurable marker is released and available for detection upon cleavage by the protease. As a result, the concentration of the measurable marker is directly related to the level of expression of the autoregulatory molecule.
机译:提供了一种用于测量活细胞内自调节分子表达的方法。在体内表达自调节分子和标记构建体,其中在翻译过程中将标记从构建体上切割下来。该方法包括表达具有通过可裂解的接头与可测量的表达标志物结合的自调控分子的构建体。可裂解的接头是蛋白酶的底物,其在翻译过程中在体内作用于其底物。翻译过程中的切割使自调控分子能够像天然形式一样正常折叠。释放可测量的标记物,并在被蛋白酶切割后可用于检测。结果,可测量标志物的浓度与自调节分子的表达水平直接相关。

著录项

  • 公开/公告号US2015010925A1

    专利类型

  • 公开/公告日2015-01-08

    原文格式PDF

  • 申请/专利权人 THE JOHNS HOPKINS UNIVERSITY;

    申请/专利号US201414317631

  • 发明设计人 ZACHARY HENSEL;JIE XIAO;

    申请日2014-06-27

  • 分类号C12Q1/37;G01N33/58;C07K14/245;

  • 国家 US

  • 入库时间 2022-08-21 15:19:31

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