通用真核表达载体的构建及其活性检测  

Construction of Universal Eukaryotic Vectors and Their Application in EPO Expression

作　　者：梅柱中[1] 岳军明[1] 张茂林[1] 乔桂林 殷震[1] 

机构地区：[1]解放军军需大学基因工程实验室,吉林长春130062

出　　处：《中国兽医学报》2000年第3期243-245,共3页Chinese Journal of Veterinary Science

基　　金：国家"8 63"计划资助项目! ( Z2 1-0 4-0 3 )

摘　　要：以质粒 pc DNA3和 p SV2 -dhfr为基础 ,构建了 2个通用表达载体 p MCD-A和 p MCD-B,然后将 EPOminigene克隆于它们的多克隆位点之中 ,得到表达载体 p AE和 p BE,经脂质体导入 CHO-dhfr- 细胞瞬时表达 ,结果表明 ,p MCD-A和 p MCD-B表达效果较好 ,表达产物经网织红细胞法测定 。To achieve high expression of heterologous gene, two universal eukaryotic vectors: pMCD A and pMCD B were constructed from pcDNA3 and pSV2 dhfr. Then EPO minigene were cloned into their multiple cloning sites. The recombinant plasmids were pAE and pBE. They were transfected into CHO dhfr - cells by lipofectin respectively. Their transient expression showed that they can express heterologous gene in CHO dhfr - cells efficiently. By reticulocyte counting, all the secreted EPO revealed their biological activity in vivo.

关 键 词：真核表达 通用载体 人促红细胞生成素 生物活性 

分 类 号：Q78[生物学—分子生物学]