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    • 2. 发明申请
    • MULTI-COLOR IN VITRO TRANSLATION
    • 多色翻译
    • WO2005017182A2
    • 2005-02-24
    • PCT/US2004/019097
    • 2004-07-21
    • THE JOHNS HOPINKS UNIVERSITY SCHOOL OF MEDICINETRAVERSO, Carlo, GiovanniDIEHL, FrankKINZLER, Kenneth, W.VOGELSTEIN, Bert
    • TRAVERSO, Carlo, GiovanniDIEHL, FrankKINZLER, Kenneth, W.VOGELSTEIN, Bert
    • C12Q
    • G01N33/6845G01N33/542
    • In vitro translation is a widely used tool for both analytical and preparative purposes. For analytical purposes, small amounts of proteins are synthesized and visualized through labeled amino acids incorporated during translation. The radioactively labeled amino acids originally used, such as [ 35 S]methionine or [ 14 C]leucine, have been superceded by the addition of antigenic tags or the incorporation of biotin-labeled or fluorescently-labeled amino acids. Such non-radioactive tags are simpler to visualize following translation and do not pose a radiation hazard. Among the non-radioactive tags, fluorescently labeled-lysine is the most convenient, as proteins that have incorporated this amino acid can be directly visualized following gel electrophoresis. Multiple fluorophores introduced into proteins significantly extend their utility, particularly for the comparison of in vitro translated proteins from related sources. This methodology can be employed for several purposes, including the simplified detection of rare truncating mutations in clinical samples from cancer patients.
    • 体外翻译是用于分析和制备目的的广泛使用的工具。 为了分析的目的,通过在翻译过程中并入的标记氨基酸合成少量蛋白质并使其可视化。 最初使用的放射性标记的氨基酸,例如[35 S]甲硫氨酸或[[14 C]亮氨酸)已经通过加入抗原标签或引入生物素标记或荧光标记的氨基酸而被替代。 这种非放射性标签更容易可视化翻译,并且不会造成辐射危害。 在非放射性标签中,荧光标记的赖氨酸是最方便的,因为结合了该氨基酸的蛋白质可以在凝胶电泳后直接显现。 引入蛋白质的多种荧光团显着扩展其效用,特别是用于比较来自相关来源的体外翻译蛋白质。 该方法可用于多种目的,包括简化检测癌症患者临床样品中罕见的截短突变。