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STAT3基因shRNA慢病毒载体的构建与鉴定

, PP. 260-262

Keywords: STAT3,shRNA,慢病毒

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Abstract:

目的构建并鉴定靶向人STAT3基因的shRNA慢病毒载体。方法针对人STAT3基因设计有效靶序列,合成相应的shRNADNA寡核苷酸链;退火形成双链DNA后,与经过AgeI和EcoRI内切酶双酶切后的pGCSIL-GFP载体连接,产生shRNA慢病毒载体。PCR筛选阳性克隆,测序鉴定。以293T细胞包装生成完整病毒颗粒并测定其滴度。结果PCR鉴定与DNA测序证实合成的含STAT3shRNA慢病毒载体寡核苷酸链插入正确,浓缩病毒悬液的滴度为3×108TU?mL-1。结论成功构建了靶向人STAT3基因的shRNA慢病毒载体。

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