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慢病毒是一种具有独特优点和巨大应用潜力的哺乳动物细胞基因转移载体,我们对慢病毒载体对不同哺乳动物细胞的基因转移及表达效率进行了平行比较研究.应用第三代重组慢病毒系统构建了携带CMV启动子-EGFP报告基因表达元件的重组慢病毒Lenti-EGFP,分别对多种不同哺乳动物细胞进行转导实验,在转导48 h后应用流式细胞仪检测报告基因在不同细胞株中的转移及表达效率.我们共使用了29种哺乳动物细胞株,包括14种人类组织细胞,5种猴组织细胞,9种鼠组织细胞,1种兔组织细胞.结果显示,重组慢病毒具有良好的基因转移能力,可有效进入多数哺乳动物细胞,对不同种属来源的细胞没有表现出特别的偏嗜性,但对贴壁培养细胞的基因转移效率明显高于对悬浮培养细胞.本研究为重组慢病毒系统的合理使用提供了基础.
Lentivirus is a mammalian cell gene transfer vector with unique advantages and great potential for application. We carried out a parallel comparative study of gene transfer and expression efficiency of lentiviral vectors on different mammalian cells.Last three-year-old recombinant lentiviral system A recombinant lentivirus Lenti-EGFP carrying the CMV promoter-EGFP reporter gene expression element was constructed and transduced in a variety of different mammalian cells. After transduction for 48 h, the reporter gene was detected by flow cytometry in different cells Strain transfer and expression efficiency of a total of 29 mammalian cell lines, including 14 kinds of human tissue cells, 5 kinds of monkey cells, 9 kinds of murine cells, 1 kinds of rabbit tissue cells.The results showed that the recombinant lentivirus Has a good gene transfer ability, can effectively enter most mammalian cells, cells of different species did not show any particular preference, but the adherent cultured cells gene transfer efficiency was significantly higher than the suspension of cultured cells. Research provides the basis for the rational use of recombinant lentiviral systems.