电转染哺乳类动物细胞DG44-CHO的方法探讨

目的：建立并验证DG44-CHO细胞快速、无血清培养体系的转染方法。方法：将绿色荧光蛋白表达质粒pEGFP-N1电转入DG44-CHO细胞，用流式细胞仪检测绿色荧光蛋白的表达量；将不同重组蛋白表达质粒C1—28／GL1／pCMV163、C1-28／GL2／pCMV163和TmHL／pCMV163分别电转入DGd4-CHO细胞，ELISA检测其培养上清中相应重组蛋白的表达。结果：280V电压电击20ms、质粒用量为20pg时，转染细胞中绿色荧光蛋白的表达量最高；同样在该条件下，培养上清中重组蛋白浓度最高。结论：上述电转染条件具有一定的适用性。

Study on the Method of Electroporation for DG44-CHO Cell

Objective To explore the optimal electrotransfection condition of Chinese hamster ovary（CHO） DG44 cells （DGd4-CHO）. Methods The vector pEGFP-N1 was used as a reporter gene which could express the green fluorescent protein（GFP）. The transfection efficiency and production of protein were identified through FACS analysis. The vector C1- 28/GL1/pCMV, C1-28/GL2/pCMV and TmHL/pCMV163 which carried the different foreign genes were also electrotransfered into DG44-CHO, and the recombinant proteins expression were tested by ELISA. Results＂ The time and pulse voltage as well as DNA quality were very important to the transfection efficiency. The optimum combination of three factors are as follows 280 V, 20 ms and 20 lag vector DNA. The expression level of the recombinant proteins demonstrated that the best condition is same to pEGFP-N1. Conclusion The results showed that the electrotransfection will be very valuable for the expression of recombination protein.