论文部分内容阅读
用电击孔(electroporation)转染方法已成功地使化学合成的红细胞生成素(EPO)基因在猿猴肾细胞(COS-7)中表达。对转染条件、表达质粒(pSVL-EPO)DNA含量及转染后不同时间表达产率的观察和研究表明:电击孔缓冲液及相应电压的选择与表达产率有关,且表达质粒DNA量在50～100μg,转染细胞存活率在50％左右时,表达效果好。在转染后24h,便可在细胞上清液中测到EPO活性,48～72h达高峰,EPO活性可达9.7U/ml。
Chemically synthesized erythropoietin (EPO) genes have been successfully expressed in simian kidney cells (COS-7) using the electroporation transfection method. The transfection conditions, the DNA content of expression plasmid (pSVL-EPO) and the expression of DNA at different times after transfection were observed and studied. The results showed that the selection of electroporation buffer and the corresponding voltage were related to the expression yield, 50 ~ 100μg, transfected cell survival rate of about 50%, the expression effect is good. At 24 h after transfection, EPO activity was detected in the cell supernatant and peaked at 48-72 h with an EPO activity of 9.7 U / ml.