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2. 发明申请

WO2012067402A9 RED FLUORESCENT PROTEIN FRAGMENT HAVING A SELF-ASSEMBLY ACTIVITY, METHOD FOR PREPARING SAME, AND METHOD FOR ANALYZING PROTEIN-PROTEIN INTERACTIONS USING SAME 审中-公开
标题翻译：具有自组装活性的红色荧光蛋白片段，其制备方法以及使用相同方法分析蛋白质 - 蛋白质相互作用的方法
公开(公告)号：WO2012067402A9
公开(公告)日：2012-08-09
申请号：PCT/KR2011008697
申请日：2011-11-15
申请人： KOREA RES INST OF BIOSCIENCE , CHUNG BONG HYUN , CHUNG YOUNG WON , KEEM JOO OAK
发明人： CHUNG BONG HYUN , CHUNG YOUNG WON , KEEM JOO OAK
IPC分类号： C07K14/435 , C12N15/12 , G01N33/52 , G01N33/68
CPC分类号： C07K14/43595 , C07K2319/00 , G01N33/533
摘要： The present invention relates to a red fluorescent protein fragment having a self-assembly activity, to a method for preparing same, and to a method for analyzing protein-protein interactions using same. The red fluorescent protein fragment (mPlum 1 E16V) according to the present invention is prepared by coupling the ß-strand at position 1 of the red fluorescent protein (mPlum) after the ß-strands at positions 2 to 11 via a flexible peptide linker so as to prepare a circularly permutated mPlum protein (CpmPlum), replacing glutamic acid (Glu) at position 16 of the amino acid sequence of the ß-strand at position 1 of the circularly permutated mPlum protein (CpmPlum) with valine (Val) so as to prepare a mutant (CpmPlum E16V), separating the mPlum 1 peptide (mPlum 1 E16V), which is mutated alone, from mPlum 2-11 protein and self-assembling the separated mPlum 1 peptide and mPlum 2-11 protein in order to emit a red fluorescent signal, thus enabling the red fluorescent protein fragment to have a self-assembly activity. Accordingly, the red fluorescent protein fragment (mPlum 1 E16V) having a self-assembly activity according to the present invention may serve as a biosensor that can be effectively used in analyzing the position of a protein in a living cell and the function of a protein such as protein-protein interaction.
摘要翻译：本发明涉及具有自组装活性的红色荧光蛋白片段，其制备方法，以及使用其的蛋白质 - 蛋白质相互作用分析方法。根据本发明的红色荧光蛋白片段（mPlum 1 E16V）通过如下制备：将β链在位置2至11之后的红色荧光蛋白（mPlum）的位置1处的β链经由柔性肽接头偶联（CpmPlum），用缬氨酸（Val）置换环化的mPlum蛋白质（CpmPlum）的1位的β链的氨基酸序列的第16位的谷氨酸（Glu），从而制备循环置换的mPlum蛋白质以制备突变体（CpmPlumE16V），将单独突变的mPlum1肽（mPlum1E16V）从mPlum2-11蛋白质中分离出来并自我组装分离的mPlum1肽和mPlum2-11蛋白质以便发射红色荧光信号，从而使红色荧光蛋白片段具有自组装活性。因此，根据本发明的具有自组装活性的红色荧光蛋白片段（mPlum 1 E16V）可以用作生物传感器，其可以有效地用于分析活细胞中蛋白质的位置和蛋白质的功能如蛋白质 - 蛋白质相互作用。

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