DNA疫苗在小鼠体内的组织分布及安全性研究

TISSUE DISTRIBUTION AND SAFETY STUDIES OF DNA VACCINE IN MICE

目的 通过检测绿色荧光蛋白基因在小鼠体内的表达 ,研究质粒 pEGFP -N3在组织分布及应用安全性。 方法 带有增强绿色荧光蛋白 (EnhanceGreenFluorescentProtein ,EGFP)基因的质粒pEGFP -N3的转化、扩增、大量提取及纯化 ,左大腿肌注BALB/c小鼠 ,注射后不同时间剖杀 ,取注射部位肌肉、心、脑、肝、脾、肾作冰冻切片 ,荧光显微镜下镜检。结果 质粒pEGFP -N3在小鼠注射部位肌肉及心内膜细胞内表达EGFP ,荧光 4 8h内最强 ,1w明显减弱 ,2w消失。其它组织及对照组无黄绿色荧光。结论 外源性EGFP基因能在小鼠细胞内表达 ,它可作为基因表达的检测标签 ,而质粒 pEGFP -N3不与机体染色体重组 ,应用安全 ,可作为DNA疫苗的理想载体。

Aim Tissue distribution and safety of pEGFP-N3 were studied by detecting the enhance green fluorescent protein (EGFP) gene expressing in mice.Methods Transformation,amplification,extraction and purification of plasmid pEGFP-N3,which contain enhance green fluorescent protein (EGFP) gene.The mice were killed at different time after the intramuscular injection of the plasmid.Prepared the slices of tissue by using their muscle (located in place of injection),heart,brain,liver,spleen and kidney.The slices were detected by using fluorescent microscope.Results The plasmid pEGFP-N3 expressed in tissues of injected muscle and endocardium.The fluorescent was stronge from 24h to 48h after transfection.It weakened in 1 week and disappeared in 2 weeks.There were no fluorescent exist in other tissues and the contrast groups.Conclusion Foreign EGFP DNA can expressed green fluorescence in tissue cells of mouse.It suggested that EGFP could make as a maker of gene expression.Plasmid pEGFP-N3 is an ideal vector for DNA vaccine because it is safe and no genetic toxicity in vivo.