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Construction and Identification of pTRE2hyg-rhEPO Tetracycline Response Eukaryotic Expression Vector

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Author:
No author available
Journal Title:
JOURNAL OF CHINESE PHYSICIAN
Issue:
9
DOI:
10.3760/cma.j.issn.1008-1372.2005.09.005
Key Word:
重组人促红细胞生成素;真核表达载体;基因扩增;序列分析

Abstract: 目的构建并鉴定含有四环素调控的pTRE顺式作用元件的质粒型载体pTRE2hyg-rhEPO.方法设计含BamH I和Cla I酶切位点的hEPO基因引物,采用RT-PCR法从含有rhEPO基因的CHO细胞中克隆hEPO基因片段,亚克隆至pTRE2hyg真核表达载体.转化感受态大肠杆菌Top10,酶切鉴定阳性重组子,并进行序列测定.结果经RT-PCR能扩增出604bp的片段,与预期片段大小相符,构建的重组体经酶切鉴定能切出插入片段,测序结果与预期序列完全一致.结论成功构建了重组人EPO四环素调控真核表达载体pTRE2hyg-rhEPO.

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