摘要
建立了以磁性纳米颗粒为载体,由电击法介导基因导入植物细胞的优化方法。制备了粒径小于10nm的磁性纳米颗粒,与绿色荧光蛋白(Green fluorescent protein,GFP)基因融合制备基因载体,同时,制备水稻悬浮细胞,加入电极杯中,调节电击条件为:电压分别是800,1000和1200V,电容25μF,电阻200Ω,直径10mm,电击数次。利用倒置荧光显微镜观察转化后的悬浮细胞,可以明显看到绿色荧光蛋白在水稻细胞内表达。说明纳米颗粒基因载体在电击作用下能有效进入细胞,利用磁性纳米颗粒作为基因载体电击法转化植物细胞初步成功。
An optimized transgenic method using magnetic nanoparticle as rice transgene vector mediated by electroporation was studied. Magnetic nanoparticles were synthesized with diameter less than 10 nm. The nanoparticles obsorbed green fluorescent protein （GFP） gene to form gene vectors, and the suspended rice cells were prepared in liquid medium of murashigs and skoog medium（MS）, and then were mixed into electrode cup. The voltage was regulated to 800, 1000 and 1200 V respectively. Under the electroporation conditions that capacitance was 25 μF and resistivity was 200 Ω, the electrode cup of 10 mm diameter was shocked for three times. Green fluorescence could be obviously observed in transformed suspended cells in inverted fluorescence microscope. This result shows that nanoparticle gene vector can be transferred into plant cells by electroporation. Therefore, magnetic nanoparticles as gene vector can successfully transfect plant cells via electroporation.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2010年第5期617-621,共5页
Chinese Journal of Analytical Chemistry
基金
国家高校“985工程”科技创新平台(No.200501)资助项目
关键词
磁性纳米颗粒
基因载体
电击
基因转导
水稻细胞
Magnetic nanoparticle
Gene vector
Electroporation
Gene transfer
Rice cell
