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首页> 外文会议>International symposium on medical and pharmaceutical biotechnology >Construction of a cellular model for detecting utrophin expression
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Construction of a cellular model for detecting utrophin expression

机译：检测尿道表达的蜂窝模型的构建

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Duchenne muscular dystrophy is caused by the absence of the muscle cytoskeletal protein dystrophin. Utrophin is an autosomal homologue of dystrophin, and overexpression of utrophin is expected to compensate for the dystrophin deficit. Our aim is to establish a convenient method for observation the expression of utrophin and a cellular model for drug screening of duchenne muscular dystrophy. Method: Obtain three fragment of utrophin gene at exon 18 and exon 19 and the sequence surround them by PCR. TETC gene (sequence contained tetracysteine motif) was inserted in 3 ' of fragment I. The target gene was constructed with pGPKneobpA-LoxP-A and fragments I, II, III, and was transformed into C2C12 cells by electroporation. To identify homologous recombinants, transfected C2C12 cells were screened by G418, FLASH staining and PCR detection. Result: Two homologous recombinated clones of C2C12 cells were obtained. Conclution: we have constructed a cellular model for detecting utrophin expression.
机译：Duchenne肌营养不良症是由于肌肉细胞骨骼蛋白营养不良蛋白缺失引起的。泛蛋白是肌营养蛋白的常染色体同源物，并且预期尿素的过表达会弥补营养不良素缺陷。我们的目的是建立一种方便的方法，用于观察乌干蛋蛋白的表达和杜氏肌营养不良药物筛选药物筛选的表达。方法：在外显子18和外显子19处获得尿道素基因的三个片段，并通过PCR围绕它们。将TETC基因（序列含有Tetracysteine MOTIF）插入碎片I中的3'。靶基因用PGPKneobpa-loxp-A和片段I，II，III构建，并通过电穿孔转化为C2C12细胞。为了鉴定同源重组剂，通过G418，闪光染色和PCR检测筛选转染的C2C12细胞。结果：获得了两个C2C12细胞的同源重组克隆。结论：我们已经构建了一种检测尿道表达的蜂窝模型。

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