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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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ggpps基因干扰腺病毒载体的构建及鉴定
, PP. 104-107
Keywords: ggpps基因,干扰,腺病毒
Abstract:
构建ggpps基因干扰腺病毒质粒载体并加以鉴定.根据ggpps基因序列设计ggpps干扰序列引物,定向克隆至穿梭载体pshuttle-h1的bglⅱ和hindⅲ位点,pmeⅰ酶切线性化的pshuttle-h1-siggpps干扰质粒,并与腺病毒载体(padeasy-1质粒)共同转化e.colibj5183感受态细菌,产生重组腺病毒载体.用pacⅰ酶切线性化的回收质粒,转染293a细胞包装腺病毒颗粒,在倒置显微镜下观察细胞cpe,用tcid50法测定病毒颗粒的浓度,并初步观察病毒感染pc12细胞对目的基因的干扰效率.经酶切鉴定、测序证实成功构建ggpps基因干扰腺病毒载体.包装的腺病毒浓缩悬液滴度为1.995×107pfu/ml.ggpps在人中具有保守性,该病毒能在人源的wrl-68细胞中成功表达,并且对ggpps基因干扰效率达70%以上.
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