葡萄糖酸化多聚赖氨酸对A549细胞转基因效率的实验研究

The Experimental Study of the Efficiency of Gluconoylated Polylysinein Transfection of Gene into A549 Cell

目的 :研究葡萄糖酸化多聚赖氨酸介导质粒pCMVβ Gal对A5 4 9细胞的转染效率 ;观察氯喹对葡萄糖酸化多聚赖氨酸转基因效率的影响。方法 :合成葡萄糖酸化多聚赖氨酸 ,将其独自和在不同浓度的氨喹作用下 ,分别介导质粒pCMVβ Gal转染A5 4 9细胞 ,用酶联免疫法定量分析 β 半乳糖苷酶的表达水平。结果 :葡萄糖酸化多聚赖氨酸单独使用未能有效介导质粒 pCMVβ Gal转染A5 4 9细胞 ,但在氯喹作用下能显著提高其介导的转基因效率。结论 :葡萄糖酸化多聚赖氨酸单独作用未能有效介导质粒 pCMVβ Gal对A5 4 9细胞的转染 ,当加入 5 0 μM、10 0 μM氯喹时 ,其转染效率则明显提高。但当其浓度为 2 0 0 μM时 ,其转染效率反而降低。

Objective: To study the effects of gluconoylated polylysine with or without chloroquine on transfection of plasmid (report gene) into 549 cell line. Methods: Gluconoylated polylysine with various concentrations of chlorquine was synthesized to mediate transfection of pCMVβ Gal into A549 cells and the expression level of β galactosidage was measured. Results: The gluconlylated polylysine alone did not mediate transfection of pCMV β Gal into A549 cells. Chloroquine enhanced the positive effect of gluconoylated polylysine in transfection of pCMVβ Gal. The highest effciency of transfection was obteined as 100 μM of chloroquine combined with gluconoylated polylysine was used. Conclusion: Although gluconoylated polylysine alone could not play any role in the transfection of pCMVβ Gal into A549 cells, it cerntainly stimulate the transfection as combined with 50 μM or 100 μM of chlorquine.