重组质粒导入真核细胞的两种方法比较

COMPARISON BETWEEN TWO METHODS OF DELIVERING GENES INTO EUKARYOTIC CELLS

目的 :比较糖化多聚赖氨酸介导和电穿孔两种方法将重组质粒导入真核细胞的优缺点。方法 :用最佳电压、电容、温度、DNA浓度进行电穿孔转染 ,或者先使重组质粒与糖化多聚赖氨酸电性结合后 ,再与 2 .2 .15细胞共同温育的方法 ,分别将pcEP4-aC重组质粒导入 2 .2 .15细胞 ,经潮霉素筛选 ,比较两种方法的优劣。结果 :两种方法都能成功地将重组质粒导入 2 .2 .15细胞 ,但糖化多聚赖氨酸介导者 ,细胞生存时间较长。结论 :糖化多聚赖氨酸导向配体具有良好的导入重组质粒进入肝细胞的功能 。

Objective: To compare the advantage or defect between glactosylated poly-L-lysine(Gal-PLL) targeting and electroporation for delivering gene into eukaryotic cells. Method: Take an optimal electroporation program or Gal-PLL targeting to deliver recombinant plasmid pcEP4-aC into 2.2.15 cells, respectively, and then to compare the advantage or defect between them. Result: Both methods could deliver pcEP4-aC into 2.2.15 cells effectively. However, after acceptance of pcEP4-aC, the 2.2.15 cells would survive longer when by Gal-PLL targeting. Conclusion: Gal-PLL targeting ligand is not only able to deliver gene into hepatocytes effectively, but is also more suitable for applying to in vivo use.