| 磷脂氢谷胱甘肽过氧化物酶的克隆及在大肠埃希菌中的表达 |
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| 引用本文： | 赵文然,周令望,刘艺,钟学宽. 磷脂氢谷胱甘肽过氧化物酶的克隆及在大肠埃希菌中的表达[J]. 中国地方病学杂志, 2004, 23(2): 119-120 |
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| 作者姓名： | 赵文然 周令望 刘艺 钟学宽 |
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| 作者单位： | 150086,哈尔滨医科大学中国疾病预防控制中心地方病控制中心克山病研究所 |
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| 基金项目： | 国家自然科学基金资助项目(30170803)，黑龙江省科技攻关资助项目(GB01C10601) |
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| 摘 要： | 目的:在大肠埃希菌中表达磷脂氢谷胱甘肽过氧化物酶(phospholipid hydroperoxide glutathione peroxidase，PHGPx)，以期用于抗PHGPx抗体的制备。方法:用特异引物从基因文库中扩增出PHGPx cDNA，与表达载体重组，转化大肠埃希菌。重组体中的PHGPx经点突变作用，编码硒半胱氨酸的密码子UGA变为编码半胱氨酸的UGU。结果:序列分析表明，克隆载体中包含PHGPx的完整编码序列及3‘未翻译区序列；经SDS-PAGE及Western Blot证实PHGPx在大肠埃希菌中获得了成功表达。结论:突变的PGPGx可以在大肠埃希菌中高效表达并可用于制备抗PHGPx抗体。
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| 关 键 词： | 磷脂氢谷胱甘肽过氧化物酶 克隆 基因表达 大肠埃希菌 点突变 抗体 |
| 修稿时间： | 2003-12-23 |
Cloning and expression of human phospholipid hydroperoxide glutathione peroxidase in Escherichia coli |
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| ZHAO Wen-ran,ZHOU Ling-wang,LIU Yi,ZHONG Xue-kuan. Cloning and expression of human phospholipid hydroperoxide glutathione peroxidase in Escherichia coli[J]. Chinese Jouranl of Endemiology, 2004, 23(2): 119-120 |
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| Authors: | ZHAO Wen-ran ZHOU Ling-wang LIU Yi ZHONG Xue-kuan |
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| Affiliation: | ZHAO Wen-ran,ZHOU Ling-wang,LIU Yi,ZHONG Xue-kuan. Institute of Keshan Disease,Chinese Reaserch Center for Endemic Disease Control,Harbin Medical University,Harbin 150086,China |
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| Abstract: | Objective To express human phospholipid hydroperoxide glutathione peroxidase(PHGPx) in Escherichia coli ( E.coli ). Methods The cDNA of PHGPx was amplified from a human testis library using specific primers and cloned into the expression vector pRSET. The selenocysteine codon UGA in PHGPx was mutated into cysteine codon UGU. E.coli was transformed by the recombinant structure. Results The entire structure gene and the 3' UTR (untranslated region ) of PHGPx were cloned into the cloning vector and verified by sequencing. Conclusion The mutated human PHGPx is expressed in E.coli and it could be used as antigen in the preparation of anti-PHGPx antibody. |
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| Keywords: | Phospholipid hydroperoxide glutathione peroxidase Clone Point mutation Expression |
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