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METHODS FOR PREPARING TRUNCATED PROTEINS FROM NMD-ESCAPE NUCLEIC ACID MOLECULES

机译:由NMD-蔗糖核酸分子制备截短蛋白的方法

摘要

A method for preparing a truncated protein from an NMD(nonsense-mediated mRNA decay)-escape nucleic acid molecule is provided to obtain the truncated protein, which is used as a tumor antigen and used for developing a dendritic cell therapeutic agent. A method for screening a compound inhibiting a nonsense-mediated translational repression(NMTR) is provided to be used in order to screen a material capable of generating the truncated protein from the NMD-escape nucleic acid molecule through a high-through screening. A method for preparing a truncated protein from an NMD-escape nucleic acid molecule comprises a step of inhibiting an NMTR in a cell by preventing an mRNA of the NMD-escape nucleic acid molecule from being coupled to a polysome, wherein the NMD unable to express the truncated protein is BCL2-associated X protein(BAX), human mutS homologue 3(hMSH3), TATA box binding protein(TBP)-associated factor 1B(TAF1B) or transforming growth factor beta receptor 2(TGFBR2). A method for screening a compound inhibiting the NMTR comprises the steps of: (a) transfecting an NMD-escape nucleic acid molecule having a premature termination codon into a cell; (b) contacting the cell with a candidate compound; and (c) detecting expression of the truncated protein from the nucleic acid molecule.
机译:提供一种从NMD(无义介导的mRNA衰变)逃逸核酸分子制备截短蛋白的方法以获得截短蛋白,其被用作肿瘤抗原并用于开发树突状细胞治疗剂。提供一种用于筛选抑制无义介导的翻译抑制(NMTR)的化合物的方法,以便通过高通量筛选来筛选能够从NMD逃逸核酸分子产生截短的蛋白质的材料。由NMD逃逸核酸分子制备截短蛋白的方法包括通过防止NMD逃逸核酸分子的mRNA与多核糖体偶联来抑制细胞中NMTR的步骤,其中NMD无法表达截短的蛋白是BCL2相关的X蛋白(BAX),人类mutS同源物3(hMSH3),TATA盒结合蛋白(TBP)相关的因子1B(TAF1B)或转化生长因子β受体2(TG​​FBR2)。筛选抑制NMTR的化合物的方法包括以下步骤:(a)将具有提前终止密码子的NMD逃逸核酸分子转染到细胞中; (b)使细胞与候选化合物接触; (c)从核酸分子检测截短的蛋白质的表达。

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