阳离子脂质体介导的虫荧光素酶基因表达

Cationic liposome mediated-transter and expression of firefly luciferase gene

目的 研究阳离子脂质体Lipofectin介导虫荧光素酶基因在不同细胞株中的表达。方法 质粒pDR2luc在大肠杆菌DH5a中扩增后用碱裂解法提取,并经Sepharose 2B凝胶过滤柱层析,通过凝胶电泳及紫外分光光度计分析纯度、定量,以Lipofectin分别转染人肝癌细胞株HepG2、SMMC7721、人肾癌细胞株GRC及非洲绿猴肾细胞COS7,然后以液体闪烁计数仪单光子计数法测定荧光素酶活性。结果 荧光素酶活性在4种细胞株中均有表达,且均有显著性差异(均P<0.05)。结论 阳离子脂质体Lipofectin可用于多种真核细胞基因转染。

Objective To study the transfer and expression of firefly lucificerase mediated by cationic liposome in four cell lines.Methods Plasmid pDR 2luc were amplified in E.coli DH 5a strain cells,extracted by alkaline lysis and purified by passage through Sepharose 2B column chromatography.Four cell lines including human hepatocarcinoma cell line HepG 2 and SMMC7721,human renal carcinoma cell line GRC and simian kidney cell line COS 7 were be transfected by cationic liposome Lipofectin.The activity of firefly lucifierase gene were detected with scintillation count.Results The activity of firefly lucificerase gene were detected in four cell lines significantly( P <0 05).Conclusion Cationic liposome Lipofectin is a kind of cytofectin capable of transferring DNA into various cells.