以外源红细胞生成素cDNA的表达产物为指标,研究了运载DNA和重组表达质粒的构象对电穿孔转染CHO细胞的效率的影响.结果250mg/L的运载DNA可使外源基因表达水平提高3倍;线性化质粒DNA比超螺旋DNA更适合于用电穿孔方法获得永久表达.这一结果提示,运载DNA的存在和质粒DNA的线性化对提高电穿孔转染CHO细胞的效率是必须的.
The effects of DNA conformation and carrier DNA on the efficiency of electroporation are studied using the expression of erythropoietin cDNA in CHO cells as an indicator. It turns out that 250mg/L carrier DNA increases the expression level by 3 fold,and linearized plasmid DNA is more suitable than supercoiled DNA to obtain stable transfection by electroporation. The results suggest that it is essential to linearize plasmid DNA and cotransfect with apporporiate carrier DNA to obtain stable and high level expression of foreign genes by means of electroporation.
卢柏松,黄培堂. DNA对电穿孔转染效率的影响[J].生物化学与生物物理进展,1995,22(3):256-260
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