氨基葡甘聚糖的质粒DNA转染载体作用的研究

目的 :发展一种新的半人工合成的质粒DNA转染载体。方法 :用氨基化方法将纯化的葡甘聚糖阳离子化 ,用凝胶阻滞检测法 (gelretardationassayforcomplexformation)观察氨基葡甘聚糖 DNA复合物的形成 ,以及用氨基葡甘聚糖作报告基因质粒pEGFP Cl的载体 ,转染HEK2 93细胞 ,观察转染效率。结果 :葡甘聚糖氨基化后溶液离子强度增大 2 0倍左右 ,氨基葡甘聚糖可与质粒DNA形成复合物 ,形成的复合物转染HEK2 93细胞后 ,报告基因pEGFP Cl获得阳性表达。氨基葡甘聚糖最大至 1 0 %仍未显示细胞毒性。结论 :氨基葡甘聚糖可发展为DNA载体系统 ,在HEK2 93细胞中获得理想的目的基因表达。

Plasmid DNA Delivery Effects Mediated by Amino-glucomannan in vitro by Using Report Genes

A semi artificial non\|viral delivery systems, amino\|glucomannan for gene transfection have been investigeted. The polysaccharides becomes cationic by amminification, then it could be aggregated into various nano\|particle upon the degree of aminification. Its ionic strength increases to 20 folds. The character of amino\|glucomannan to combine a report gene, plasmid pEGFP\|Cl and the transfection effects of amino\|glucomannan/DNA complex to HEK293 cell line were explored. The results showed that the amino\|glucomannan is a non\|specific plasmid DNA delivery carrier with less cytotoxicity, so it should receive much attention as a novel gene carrier recently.