慢病毒介导绿色荧光蛋白基因转染大鼠软骨细胞表达的研究Expression of green fluorescent protein gene in rat chondrocytes mediated by lentiviral vector
陈文雄;杨小红;李斯明;
摘要(Abstract)：
[目的]构建携带绿色荧光蛋白基因(green fluorescent protein,GFP)的慢病毒载体系统,通过感染大鼠软骨细胞,观察其感染能力及GFP的表达情况。[方法]以载体质粒pLentiLox3.7、包装质粒pRSV-REV、pMDL g/p RRE及包膜质粒VSV-G共转染293T细胞制备慢病毒。分离、培养大鼠关节软骨细胞,将制备好的慢病毒颗粒感染软骨细胞,在荧光显微镜下观察报告基因GFP的表达情况。[结果]重组慢病毒载体系统转染293T细胞96 h后可观测到较强的绿色荧光蛋白的表达,收集上清测得病毒滴度约为3.0×103ifu/μl,并且在80%大鼠软骨细胞中观察到GFP的表达。[结论]成功构建了携带GFP的重组慢病毒载体系统,对大鼠关节软骨细胞转染效果良好,为将来结合RNA干扰技术对软骨细胞进行基因改造提供了基础。
关键词(KeyWords)： 慢病毒载体;绿色荧光蛋白;软骨细胞
基金项目(Foundation):
作者(Author): 陈文雄;杨小红;李斯明;
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DOI:
参考文献(References)：
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