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    • 6. 发明申请
    • METHOD FOR SELECTIVE LABELING OF PROTEIN PRODUCED BY IN VITRO TRANSLATION BY USING A MARKER FROM TRNA PREPARED BY USING IN VITRO TRANSCRIPTION
    • 通过使用通过使用体外转录制备的TRNA的标记通过体外翻译产生的蛋白质的选择性标记的方法
    • US20110014714A1
    • 2011-01-20
    • US12812382
    • 2009-01-07
    • Seongjun YoonSooYoun JunSangHyeon Kang
    • Seongjun YoonSooYoun JunSangHyeon Kang
    • G01N33/68C12P19/34C07H21/02
    • G01N33/533G01N33/532
    • The present invention relates to a method for the preparation of a labeling material (labeling agent) usable for non-radioactive selective labeling of proteins produced by in vitro translation with fluorophore or biotin; and a method for selective labeling of proteins produced by in vitro translation using the said labeling material. More specifically, the present invention provides a method for preparing a labeling material usable for selective labeling of a target protein produced only from the gene added to the cell-free protein synthesis reaction mixture by an experimenter without labeling pre-existing proteins in the reaction mixture, for which only one type of tRNA prepared by in vitro transcription is used instead of “total tRNA mixture”. The present invention also provides a method for labeling of the protein produced by in vitro translation by using the labeling material prepared by the above method. According to the present invention, the preparation of a labeling material becomes easier and the protein labeled with the labeling material of the present invention provides much improved signals than the protein labeled with the labeling material of the conventional method prepared by using “total tRNA mixture” as a tRNA material.
    • 本发明涉及制备可用于通过用荧光团或生物素的体外翻译产生的蛋白质的非放射性选择性标记的标记材料(标记试剂)的方法; 以及使用所述标记材料通过体外翻译产生的蛋白质选择性标记的方法。 更具体地说,本发明提供了一种可用于选择性标记仅通过实验者从无基因蛋白质合成反应混合物中添加的基因而产生的靶蛋白的标记物质的方法,而不在反应混合物中标记预先存在的蛋白质 ,其中仅使用通过体外转录制备的一种类型的tRNA代替“总tRNA混合物”。 本发明还提供了通过使用通过上述方法制备的标签材料通过体外翻译产生的蛋白质的标记方法。 根据本发明,标记材料的制备变得更容易,并且用本发明的标记材料标记的蛋白提供比使用通过使用“总tRNA混合物”制备的常规方法的标记材料标记的蛋白质更大的改善的信号, 作为tRNA材料。