红细胞生成素(EPO)在中国仓鼠卵巢细胞(CHO)中稳定表达  

Stable Expression of Human Erythropoietin cDNA in Chinese Hamster Overy(CHO) Ce lls

作　　者：赵玮[1] 戴长柏[1] 孙茂盛[1] 

机构地区：[1]中国医学科学院中国协和医科大学医学生物学研究所

出　　处：《云南大学学报（自然科学版）》1999年第3期175-178,共4页Journal of Yunnan University(Natural Sciences Edition)

摘　　要：将ＥＰＯｃＤＮＡ分别插入真核表达质粒载体ｐＳＶ２ｄｈｆｒ和ｐｃＤＮＡ３的适当位点，用电脉冲法导入ＣＨＯ细胞，用ＥＬＩＳＡ法检测ＥＰＯ的表达量，用ＴＦ１细胞检测ＥＰＯ表达产物的生物活性．结果表明，用ｐＳＶ２ｄｈｆｒ作为载体，在抗ＭＴＸ阳性细胞克隆中，获得了表达ＥＰＯ（２５８ｎｇ／ｍＬ培养液）的ＣＨＯ细胞株，并且表达产物具有生物活性．进一步的传代和冻存试验表明。Expression plasmid pSV2 dhfr/EPO and pcDN A3/EPO were constructed and electroporated into CHO dhfr cells.The result s of EPO ELISA test showed that pSV2 dhfr/EPO can express EPO in CHO cslls while pcDNA3/EPO can not.Cell line with stable rhEPO expression were obtained which secreted rhE PO at the level of 258?ng·mL -1 culture supernate.And it was proved that the recombinant EPO had biological activity.

关 键 词：红细胞生成素 CHO细胞 真核表达 卵巢细胞 

分 类 号：R973.3[医药卫生—药品] Q78[医药卫生—药学]