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Pri-miRNA21/23a重组病毒载体的构建及其在肝细胞中的表达鉴定

Letters in Biotechnology(2017)

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Abstract
目的:构建Pri-miRNA-21/23A基因的重组病毒载体,并在L-02肝细胞中获得表达.方法:设计并合成PrimiRNA-21/23a的基因产物,插入含绿色荧光蛋白Zsgreen基因的真核表达载体pCI MammaLian中,以重组质粒为模板,设计扩增含Zsgreen基因的Pri-miRNA序列产物,并将其与pCDH-CMV-MCS-EF 1-Puro病毒载体连接,将PrimiRNA重组病毒载体转染293T细胞后进行病毒包装,病毒上清转染L-02肝细胞后用荧光显微镜及实时荧光定量PCR确认转染效果.结果:荧光定量PCR结果显示重组病毒上清转染L-02肝细胞感染效果良好,经荧光显微镜观察,证实重组病毒载体能在细胞中表达蛋白.结论:构建了Pri-miRNA21/23a重组病毒表达载体并在L-02肝细胞中表达,奠定了miRNA进一步功能研究的基础.
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Key words
microRNA,miR-21,miR-23a,lentivirus vector
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