【摘要】 盘基网柄菌(Dictyostelium discoideum)是一个应用广泛的模式生物,非常适合用来研究胞质分裂、细胞运动、吞噬作用、趋化性、趋电性、信号转导以及个体发育过程中的细胞分化。该实验主要介绍电穿孔技术转化绿色荧光蛋白标记肌动蛋白基因质粒(Lifeact-GFP)进入盘基网柄菌活细胞中,抗性筛选(潮霉素B)获得阳性克隆子,最后借助荧光显微镜观察绿色荧光蛋白标记的微丝在盘基网柄菌的分布情况。综合性实验训练可提高学生的学习兴趣和综合运用理论知识的能力,进而可培养学生的科学研究思维。更多还原

【Abstract】 Dictyostelium discoideum is a powerful model organism that is ideal for studying cytokinesis,cell movement,phagocytosis,chemotaxis,electrotaxis,signal transduction,and cell differentiation.This experiment describes the use of electroporation technology to transform the Lifeact-GFP (Lifeact-green fluorescent protein) into living cells of Dictyostelium discoideum,then screening for positive clones with resistance to hygromycin B.The Lifeact-GFP-labeled microfilaments distributed in Dictyostelium discoideum are observed by fluorescence microscopy.Through comprehensive laboratory course training,the interest of learning and the ability ofapplying to theoretical knowledge of undergraduate students can be improved,thereby training the students in scientific thinking.更多还原