慢病毒包装体系的优化及其感染马胚胎成纤维细胞的研究

Optimization of lentiviral packaging system and study on infection of equine fibroblasts

为了建立高效率的病毒包装体系,完善研究蒙古马相关基因功能的基础手段,试验采用脂质体3000和脂质体LTX两种试剂对编码绿色荧光蛋白基因的慢病毒质粒的转染效率和慢病毒感染效率进行比较,并使用其中转染效率和感染效率较优的脂质体进一步感染马胚胎成纤维细胞。结果表明:脂质体3000与脂质体LTX对编码绿色荧光蛋白基因的慢病毒质粒转染效率分别为(79.4±6.3)%和(74.6±6.2)%;脂质体3000与脂质体LTX包装病毒液后感染293T细胞的感染效率分别为(74.0±3.6)%和(69.2±3.4)%。两者之间的转染效率及感染效率均差异不显著(P>0.05),都可以有效改善病毒的感染,其中脂质体3000的应用效果更好,脂质体3000可成功感染马胚胎成纤维细胞,感染效率为(69.4±6.4)%。说明脂质体3000可以高效率包装慢病毒,并可用于感染马胚胎成纤维细胞。

To establish a high-efficiency virus packaging system and improve the basic means of studying the functions of Mongolian horse-related genes,two reagents,liposome 3000 and liposome LTX,were used for transfection efficiency and infection efficiency of lentiviral plasmid encoding green fluorescent protein gene.The liposomes with better transfection and infection efficiency were used to further infect the equine fibroblasts.The results showed that the transfection efficiency of Lipofectamine 3000 and Lipofectamine LTX on lentiviral vector encoding green fluorescent protein gene was(79.4±6.3)%and(74.6±6.2)%,respectively.The infection efficiency of 293T cells infected by Lipofectamine 3000 and Lipofectamine LTX was(74.0±3.6)%and(69.2±3.4)%,respectively.There was no significant difference in transfection efficiency and infection efficiency between the two groups(P>0.05),both of which could effectively improve the virus infection.The application effect of liposome 3000 was better,and the infection efficiency of liposome 3000 was(69.4±6.4)%.It indicated that Lipofectamine 3000 could efficiently package lentivirus and infect equine embryonic fibroblasts.