脂质体介导的cho细胞染——脂质体介导的基因真核细胞转染毕业论文.docVIP

脂质体介导的CHO细胞转染 ——脂质体介导的基因真核细胞转染 专业：生物工程专业 班级：2008级二班 姓名： 摘要 CHO细胞是中国仓鼠卵巢细胞，是目前生物工程上广泛使用的细胞系。真核细胞中CHO细胞是目前重组糖基蛋白生产的首选体系。 外源基因进入细胞主要有四种方法：电击法、磷酸钙法和脂质体介导法和病毒介导法。 脂质体是磷脂分散在水中时形成的脂质双分子层，又称为人工生物膜。根据膜的融合及内吞作用，可用作外源物质进入细胞的载体。 在对生物大分子的传递过程中，脂质体是十分有效的载体，无论对微生物还是对植物细胞或对哺乳动物细胞导入外源DNA，脂质体的包裹都能对抗核酸酶的消化。 脂质体的制备主要有超声波法、磷脂酰丝氨酸Ca2+离子融合法和逆相蒸发等，用超声波法可制备的单层或多层脂质小体能向细胞内引入的物质量少，故常用后两种方法制备大的脂质体。脂质体介导转染法的改进方法包括促进转染的脂类、增加转染效率的添加剂、DNA与脂质体最佳比率的确定。 本实验将携带绵羊P基因的绿色荧光蛋白真核表达载体转入中国仓鼠卵巢细胞，得到稳定表达绵羊PrP基因的CHO细胞系。得出外源导入CHO的绵羊PrP可以在CHO中稳定表达的结论。 关键词：CHO细胞、脂质体、转染 Abstract CHO cells of Chinese hamster ovary cells, biological engineering is currently widely used cell lines. Eukaryotic cells in CHO cells is currently restructuring glycosyl protein of choice for the production system. Exogenous genes into cells mainly has four kinds of methods: electric shock method, calcium phosphate and mediated by liposome and virus mediated method. Liposome is a phospholipid dispersed in water is formed when the lipid bilayer, also known as the artificial biological membrane. According to the membrane fusion and endocytosis, can be used as exogenous substances into the cell carrier. On biological macromolecules in the transfer process, liposomes are very effective carrier, regardless of the microorganism or plant cell or mammalian cells for introducing exogenous DNA, liposome encapsulated can against nuclease digestion. Preparation of liposomes are ultrasonic method, phosphatidylserine Ca2+ ion fusion method and reverse phase evaporation, using ultrasonic method to prepare monolayer or multilayer lipid globules to intracellular introduced mass is less, so commonly used two methods for preparation of large liposomes. Liposome mediated transfection method improvement methods include promoting transfection lipids, increase in transfection efficiency additives, DNA and liposome best rate determine. This experiment will carry sheep P gene green fluorescent protein eukaryotic expression vector was transfected into Chines