摘要
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 vector was widely used as gene expressing vector in mammalian cells. Yet few comparing transduction and expression efficiencies of a recombinant lentivirus in different mammalian cells were performed. In this study,we applied the 3rd generation lentivirus expression system to construct a recombinant lentivirus which was expressing EGFP as a report gene, then used it to transduce several different mammalian cell lines under the same conditions. After 48 h post of infection,the flow cytometer was employed for analysis of the transduction and expression efficiencies of EGFP gene in different cell lines. Twenty nine different cell lines were examined,including 14 human cell lines,5 monkey cell lines,9 rodent cell lines and 1 rabbit cell line. Results showed that recombinant lentivirus Lenti-EGFP was able to transduce most of the mammalian cells, preferably for the primate cell lines, and efficiencies were higher in adherent culture cell lines than in suspend culture cell lines. There no apparent partiality was observed for cells of different organ origin. This study offered essential knowledge for rationally propagating recombinant lentivirus.
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
2008年第3期392-396,共5页
Journal of Xiamen University：Natural Science
基金
教育部科学技术研究重大项目培育基金(705031)
福建省自然科学基金(C0710041)
厦门市病毒性疾病新药研发平台建设基金(3502Z20041008)资助
