一种非病毒载体的组成与体外转基因能力的研究

目的：合成羟丙基β-环糊精偶联小分子量聚乙烯亚胺的共聚物,探讨不同聚乙烯亚胺偶联率的共聚物缩合DNA的能力,探讨不同偶联率的阳离子共聚物组成与转基因效率的关系。方法：通过NMR表征,凝胶阻滞DNA试验考察其缩合DNA能力,以MTT法检测不同偶联率共聚物的细胞毒性。以不同的偶联率共聚物为载体,在不同的N/P比值条件下,将荧光素酶基因导入非洲绿猴肾细胞COS-7,通过荧光素酶的表达检测比较不同偶联率的共聚物转染效率。结果：合成的不同偶联率的共聚物缩合DNA能力不同,转染效率也不同。结论：聚乙烯亚胺偶联率较高的阳离子共聚物缩合DNA的能力较强,转染效率较高。

Study on the Composition of Copolymer and Its Gene Delivery as a Non-viral Vector in vitro

Objective： Low molecular weight polyethylenimine was coupled to HP β-cyclodextrin as copolymer for gene delivery. It was investigated the relationship between the coupling rate and the capacity of condensing DNA, the relationship between the coupling rate and the transfection efficiency of copolymer in vitro Method： The structure and the coupling rate of copolymer were measured by ^1H-NMR. The capacity of condensing DNA of copolymer was investigated by agarose gel electrphoresis assay.The cytotoxicity of copolymer was evaluated by MTT assay.The transfection efficiency of copolymer was evaluated by luciferse assay Results：The copolymer of different coupling rate showed different capacity of condensing DNA and different transfection efficiency. Conclusion： The higher coupling rate of copolymer showed higher capacity of condensing DNA and higher transfecion efficiency as gene delivery vector. It was potential candidate non--viral vector for gene therapy.