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目的 为基因免疫和基因治疗寻找一种简单、经济、有效的 DNA投递系统。方法 采用逆向蒸发法 ,用卵磷脂、胆固醇、十八胺制备成阳离子脂质体包封含绿色荧光蛋白基因的真核表达质粒 p EGFPC1,对三种不同DNA加入量所形成的 DNA/脂质体复合物的微球形、包封率、电荷性、转染真核细胞的效果以及对细胞的毒性作用进行初步检测研究。结果 所获得的 DNA/脂质体复合物呈球形、平均粒径为 5 6 2 nm;包封率达 5 5 %～ 6 5 % ,为阳离子脂质体 ;都能有效转染真核细胞 ,但转染效率有差异 ,DNA/脂质体混悬液中 DNA含量、混悬液的加入量以及细胞的作用时间 ,都对细胞毒性有不同程度的影响。结论 该方法可作为提高基因免疫效果的简单、经济、有效的手段之一
Objective To find a simple, economical and effective DNA delivery system for gene immunization and gene therapy. Methods The retroviral method was used to encapsulate the eukaryotic expression plasmid p EGFPC1 containing green fluorescent protein gene with cationic liposomes using lecithin, cholesterol and octadecylamine. DNA / lipids of three different DNA addition levels The microsphere shape, encapsulation efficiency, chargeability, the effect of transfection of eukaryotic cells and the cytotoxicity on the cells were studied preliminarily. Results The DNA / liposome complexes obtained were spherical with the average particle size of 562 nm and the encapsulation efficiency of 5- 5% ~ 65%, which were cationic liposomes. All of them could efficiently transfect eukaryotic cells, However, there are differences in transfection efficiency, DNA content in DNA / liposome suspension, the amount of suspension added and the action time of cells, have different degrees of cytotoxicity. Conclusion This method can be used as one of the simple, economical and effective means to improve the effect of genetic immunization