重组人促红细胞生成素在COS-7细胞内表达的研究

Preliminary S1tudy on Recombinant Human Erythropoietin Expressed in COS 7

目的：研究不同真核载体对人促红细胞生成素ｃＤＮＡ的表达效率。方法：人促红细胞生成素ｃＤＮＡ分别用真核表达载体ｐｃＤＮＡ３、ｐｃＤＩ、ｐＡｄＣＩ进行重组，脂质体法转化ＣＯＳ７细胞，以ＴＦ１细胞检测ＥＰＯ的表达活性。结果：感染ｐｃＤＮＡ３ＥＰＯ，ｐｃＤＩＥＰＯ，和ｐＡｄＣＩＥＰＯ的ＣＯＳ７细胞培养上清中人促红细胞生成素活性分别为０５×１０２，１５×１０３和２２×１０２。结论：已构建成可表达具生物活性的重组人促红细胞生成素的真核表达克隆。

AIM:study on expression level of recombinant erythropoietin in COS 7 using different eukaryotic expressing vector.METHODS:the human erythropoietin cDNA was inserted to eukaryotic vector pcDNA3,pcDI,and pAdCI,transfected COS 7 using Lipofectin, and detected the expressing activity level of erythrpopietin using MTT technique with TF 1. RESULTS:the supernatant of COS 7 transfected with pcDNA3 EPO,and pAdCI EPO were 0.5×10 2,1.5×10 3,and 2.2×10 2 respectively. CONCULUTION:We have constructed the recombinant clones that expressed human erythropoietin, and the chimeric intron enhanced expression of erythropoietin.