人血小板生成素(hTPO)全长 cDNA 克隆及在 CHO 细胞中的表达  

Cloning and Expression of a Full Length cDNA of Human Thrombopoietin (hTPO)in CHO Cells

作　　者：邢桂春[1] 郭树华 胡志远 张咏[1] 董波[1] 贺福初[1] 

机构地区：[1]北京放射医学研究所

出　　处：《生物工程学报》1997年第4期410-414,共5页Chinese Journal of Biotechnology

基　　金：863计划;自然科学基金

摘　　要：以人胎肝ｍＲＮＡ为模板，采用ＲＴＰＣＲ扩增了人ＴＰＯ编码区全长ｃＤＮＡ，全序列测定结果表明与国外报道序列一致；进而构建了ｐｃＤＮＡ３ＴＰＯ永久表达载体，转染ＣＨＯ细胞后经Ｇ４１８加压筛选、ＴＰＯ表达稳定性等项指标测试，获得了稳定分泌ＴＰＯ的工程细胞株。A full length cDNA of the coding region of hTPO,whose sequence is identical to those previously reported,has been amplified by means of RT PCR with human fetal liver mRNA as the templates and then sequenced.Further,this insert was subcloned into a mammalian expression vector pcDNA3 (pcDNA3 TPO),transformed into CHO cells, and screened by G418 and then tested by expression level of hTPO.A group of stably secreting engineered strains of hTPO has been obtained.

关 键 词：人血小板生成素 药物 CHO细胞 表达 CDNA 克隆 

分 类 号：R392－33[医药卫生—免疫学] R394.8[医药卫生—基础医学]