| 转译优化对EPO基因在COS7细胞中表达的影响 |
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| 引用本文： | 童涌,周向军.转译优化对EPO基因在COS7细胞中表达的影响[J].生物工程学报,1994,10(4):342-345. |
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| 作者姓名： | 童涌 周向军 |
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| 作者单位： | 第二军医大学医学生物技术和分子遗传研究所，第二军医大学医学生物技术和分子遗传研究所，第二军医大学医学生物技术和分子遗传研究所，第二军医大学医学生物技术和分子遗传研究所，第二军医大学医学生物技术和分子遗传研究所 上海 200433，上海 200433，上海 200433，上海 200433，上海 200433 |
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| 摘 要： | 利用ＣＯＳ７细胞暂时表达系统，研究转译起始序列对ＥＰＯ－ｃＤＮＡ表达的影响。通过ＤＮＡ重组技术，构建了原ＥＰＯ－ｃＤＮＡ表达载体ｐＣＳＶ－ＥＰＯ（１），其转译起始序列为５＇ＡＡＴＴＣＡＴＧＧ３＇。同时通过定点突变技术，将起始序列改变成５＇ＣＣＡＣＣＡＴＧＧ３＇，而构建了另一表达载体ＰＣＳＶ－ＥＰＯ（２）。后经序列分析证明无误后和前均通过ＤＥＡＥ－ｄｅｘｔｒａｎ法转染ＣＯＳ７细胞上清，测定结果为
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| 关 键 词： | 促红细胞生成素 基因表达 COS7细胞 |
Effect of Optimized Translational Initiation on EPO cDNA Expression in COS7 Cells |
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| Tong Yong Zhou Xiangjun Cao Yunxu Wen Zhaorong Lu Deru.Effect of Optimized Translational Initiation on EPO cDNA Expression in COS7 Cells[J].Chinese Journal of Biotechnology,1994,10(4):342-345. |
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| Authors: | Tong Yong Zhou Xiangjun Cao Yunxu Wen Zhaorong Lu Deru |
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| Abstract: | Using DNA recombination technique, we constructed a EPO-cDNA expression vector pCSV-EPO (1), its sequence around the ATG initiator codon was 5' AATTCATGG 3'. By applying site-directed mutagenesis to the EPO-cDNA, another EPO-cDNA expression vector was constructed, mutations were created near the ATG initiator codon, its sequence was changed to 5' CCACCATGG 3'. It was demonstrated by DNA sequencing. After that, the two expression vectors were introduced into COS7 cells by DEAE-dextran-mediated transfection. The EPO concentration in culture was determined by EPO-ELISA kit. The results showed that the pCSV-EPO (2) had a higher EPO expression level than the pCSV-EPO (1) had. |
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| Keywords: | Erythropoietin (EPO) site-directed mutagenesis translational initiation DNA sequencing COS7 cell |
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