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gou jian xie dai nurr1 ji yin de man bing du biao da zai ti ji zhuan ran ren qi dai jian chong zhi gan xi bao de shi yan yan jiu
Author(s): 
Pages: 35-38
Year: Issue:  2
Journal: Acta Academiae Medicinae Jiangxi

Keyword:  核受体相关因子-1 基因人脐带间充质干细胞慢病毒表达载体;
Abstract: 目的:构建携带核受体相关因子-1(nuclear receptor related factor 1,nurr1)基因的慢病毒表达载体,并测定其对人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs)的感染效率。方法使用PCR 技术克隆目的 nurr1基因,将 nurr1基因重组到慢病毒表达载体,筛选阳性克隆,将携带 nurr1基因的质粒用脂质体介导法转染入293T 细胞,获得携带 nurr1基因的重组慢病毒,再用重组慢病毒感染 hUCMSCs,通过荧光显微镜及流式细胞术检测其感染效率。结果成功构建了携带 nurr1基因的重组慢病毒载体,并获得病毒滴度为2×108 TU·mL-1的慢病毒浓缩液。成功转染重组慢病毒的 hUCMSCs 可表达绿色荧光蛋白,当病毒感染复数(mul-tiple of infection,MOI)值为60时转染效率最高为90.8%。结论携带 nurr1基因的慢病毒载体成功构建并成功转染入 hUCMSCs,当 MOI 值为60时转染效率最高。
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