基于壳聚糖纳米颗粒的基因枪法转化洋葱细胞研究  

Chitosan Nanoparticle as Gene Vehicle Mediated by Particle Bombardment in Onion Cell Transformation

作　　者：王凤华[1] 刘俊[1,2] 唐冬英[1] 薛昌刚[1] 童春义[1] 吴宪远[1] 刘选明[1] 

机构地区：[1]湖南大学生命科学与技术研究院,湖南长沙410082 [2]湖南大学物理与微电子科学学院,湖南长沙410082

出　　处：《湖南大学学报（自然科学版）》2009年第5期67-70,共4页Journal of Hunan University:Natural Sciences

基　　金：高校国家'985工程'科技创新平台资助项目(200501)

摘　　要：以交联法制备壳聚糖纳米颗粒,用透射电子显微镜和Zeta电位仪对壳聚糖纳米颗粒进行表征,发现颗粒呈球形,粒径约为50 nm,微球表面光滑、球形圆整、颗粒比较均匀、分散性好,电位约为11.1 mv;琼脂糖凝胶电泳结果显示,壳聚糖纳米颗粒能有效地结合质粒DNA,并能保护所结合的DNA防止DNase I的酶切;将含GFP基因的壳聚糖纳米颗粒复合物使用基因枪转化洋葱表皮细胞,在倒置荧光显微镜下观察,发现细胞表达了绿色荧光蛋白,表达效率为8%.Chitosan nanoparticles were synthesized with cocervation process. The particles were characterized with transmission electron microscope and Zeta-Sizer measurement. Chitosan nanoparticles were shown to be spherical structure distributed evenly, the average sizes of the particles were about 50 nm in diameter, and the particles were 11.1 mv charge. Plasmid DNA was absorbed to chitosan nanoparticles by electrostatic forces. Agarose gel electrophoresis showed that the pEGAD plasmid DNA was effectively absorbed to chitosan nanoparticles. Furthermore, the complexes could protect the absorbed DNA from DNase Ⅰ damage. Chitosan nanoparticles-DNA mixed was transformed into the onion cell via particle bombardment, and onion cells were observed under invert-fluorescence-microscope. The results showed that transformed onions with GFP gene were obtained by particle bombardment successfully. Analysis showed that the reporter GFP gene was expressed with high efficiency （8 %）.

关 键 词：纳米颗粒 基因枪 转化 洋葱细胞 

分 类 号：O613.52[理学—无机化学]