【摘要】 以我国重要水产养殖动物中国明对虾(Fenneropenaeus chinensis)贴壁培养和悬浮培养的血细胞、植块培养的淋巴(Oka)器官细胞和卵巢细胞为材料,通过磷酸钙共沉淀、脂质体转染和电穿孔等多种方法进行了导入EGFP基因的实验。结果表明,三种方法优化条件后均可将外源基因导入培养细胞,但只有通过脂质体介导的转染可以使EGFP基因得到表达。更多还原

【Abstract】 The development of the technology related to introducing the eukaryotic expression plasmid into the primarily cultured cells from Chinese shrimp (Fenneropenaeus chinensis), one of the most commercially important aquaculture marine invertebrates in China was introduced. The primary cell cultures included the adherent or suspension cultures from the haemocytes and the cell adherent cultures from the lymphoid (Oka)organ and ovary of the shrimp. The methods of gene transfer included calcium phosphate mediated transfection, liposome mediated transfection(lipofection)and electroporation. Lipofectin could be used to introduce the plasmid into the cultured cells and to express the reporter gene, EGFP(enhanced green fluorescence protein)gene, in the haemocyte suspension cultures and the cell adherent cultures from the Oka organ and ovary. The aim is to facilitate the study of shrimp immunology and transgenic studies by developing a primary culture system of the shrimp.更多还原