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Per2-shRNA慢病毒构建及嘌呤霉素筛选Per2低表达的细胞株

, PP. 252-255

Keywords: Period2基因,shRNA,U343细胞,慢病毒

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Abstract:

目的构建人Period2(Per2)基因shRNA慢病毒。方法设计并合成3对特异性人Per2基因shRNA靶序列,构建到pENTRTM/U6(GV112)慢病毒载体中,包装慢病毒,用real-timePCR筛选取最佳片段。对感染的U343细胞,经嘌呤霉素初步筛选,Westernblot鉴定Per2蛋白表达,得到Per2低表达的细胞。结果成功构建了具有Per2沉默效应的shRNA慢病毒。结论此次包装的慢病毒可以成功抑制Per2基因在胶质瘤U343细胞中的表达,并可用嘌呤霉素筛选低表达细胞株。

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