摘要
目的 比较传统的脂质体转染技术与NucleofectorTM转染技术将pLXSN CTLA4Ig TRES2 EGFP重组质粒转染入PA3 17细胞的效率。方法 分别用脂质体技术与NucleofectorTM 转染技术将pLXSN CTLA4Ig IRES2 EGFP重组质粒转染PA3 17细胞 ,应用倒置荧光显微镜、流式细胞仪检测转染后PA3 17细胞绿色荧光蛋白表达情况及阳性细胞率 ,比较两种方法的转染效率。结果 脂质体转染后 4、8h未观察到阳性细胞 ,转染后 12h阳性率为 ( 0 5 9± 0 2 ) %,转染后 2 4h阳性率为 ( 4 3 9± 2 43 ) %;NucleofectorTM 转染技术转染后约 4h观察到阳性细胞 ,转染后 12h阳性率为 ( 12 2 7± 3 3 8) %,转染后 2 4h阳性率为 ( 5 0 16± 2 41) %。两者比较 ,在转染后 12、2 4h ,后者阴性率显著高于前者 (P <0 0 1)。结论 NucleofectorTM 技术与传统的脂质体技术比较 ,是一种具有转染速度快。
Objective To compare the traditional transfection efficiency of liposome transfection technique and Nucleofector TM technique for the transfection of PLXSN CTLA4Ig TRES2 EGFP recombinant plasmid into PA317 cells. Methods Liposome transfection technique and Nucleofector TM technique were employed for the transfection of pLXSN CTLA4Ig TRES2 EGFP recombinant plasmid into PA317 cells, respectively. The expression of green fluorescent protein (EGFP) by PA317 cells and the positive cell rates were detected with inverted fluorescence microscope and flow cytometer for the comparison of the transfection efficiency by the two methods. Results Positive cells were not observed at 8 h after liposome transfection. Liposome transfection technique resulted in positive cell rates of (0 59±0 2)% and (4 39±2 43)% at 12 h and 24 h, respectively. However, Nucleofector TM technique revealed positive cells at 4 h, and resulted in positive cell rates of (12 27±3 38)% and (50 16±2 41)% at 12 h and 24 h, respectively. Conclusion Compared with the traditional liposome transfection technique, Nucleofector TM technique is a transfection technique with faster speed and higher efficiency.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2004年第2期104-106,共3页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目 ( 30 30 0 0 90 )~~
