摘要
于鸡胚19期分离生殖嵴原始生殖细胞(PGCs),用于体外培养和传代扩增;对其第2代PGCs进行组织化学法鉴定,采用3种转染方法以增强型绿色荧光蛋白转染PGCs,并对电穿孔方法转染条件进行优化,荧光显微镜下计数细胞,分析PGCs转染效率,探讨有效的转染方法及其优化条件。结果表明:电穿孔方法可获得较高的转染效率,其最优化条件:电场强度为280 V,时间常数为60μs,质粒浓度为15μg.mL-1,转染后室温静置10 min。
Isolated PGCs from chicken embryo at stage 19 for cultured and subcultured in vitro was indentified by histochemistry methods and transfected with three methods, and the optimization of electroporation parameters was emphasized. The transfection efficiencies were checked by cell conduction under fluorescent microscopy. The results were as follows： The electroporation could transfect PGCs efficiently. The most optimized parameter was achieved under the condition of voltage 280 V, constant time value 60μs, plasmid concentration 15 μg · mL^-1, and room temperature.
出处
《扬州大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2007年第3期37-41,共5页
Journal of Yangzhou University：Agricultural and Life Science Edition
基金
国家自然科学基金资助项目(30170678)
关键词
鸡胚胎
原始生殖细胞
增强型绿色荧光蛋白
转染
chicken embryo
primordial germ cells
enhanced green fluorescent
transfection
