| 辣根过氧化物酶同功酶C3基因在毕赤酵母中的表达 |
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| 引用本文： | 邵金辉,朱有名,韩金祥,吴围屏,吴坚美.辣根过氧化物酶同功酶C3基因在毕赤酵母中的表达[J].生物技术,2005,15(6):8-10. |
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| 作者姓名： | 邵金辉 朱有名 韩金祥 吴围屏 吴坚美 |
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| 作者单位： | 山东省医药生物技术研究中心,国家卫生部生物技术药物重点实验室,山东,济南,250062 |
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| 基金项目： | 山东省自然基金资助项目（“辣根过氧化物酶与丙型肝炎病毒核心蛋自的融合表达研究”,Y2004C33）. |
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| 摘 要： | 目的：克隆与表达辣根过氧化物酶同功酶C3（HRPC3）基因。方法：用PCR方法从辣根的总DNA中，扩增得到一种辣根过氧化物酶同功酶C3基因，将PCR产物连接到pMD18-T载体上，测序证明正确后，再将目的基因正确插入到巴斯德毕赤酶母表达载体pPIC9K中，含有目的基因的重组pPIC9K质粒在毕赤酶母中进行表达与分析。结果：应用毕赤酶母作为受体菌，成功表达了HRPC3基因，其活性为56IU/L。结论：毕赤酶母直接表达出了具有生物活性的辣根过氧化物酶同功酶C3。
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| 关 键 词： | 辣根过氧化物酶 基因 聚合酶链反应 巴斯德毕赤酵母 |
| 文章编号： | 1004-311X（2005）06-0008-03 |
| 收稿时间： | 2005-08-25 |
| 修稿时间： | 2005-08-252005-09-17 |
Expression of Horseradish Peroxidase Isozyme C3 Gene in Pichia pastoris |
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| SHAO Jin-hui,ZHU You-ming,HAN Jin-xiang,WU Wei-ping,WU Jian-mei.Expression of Horseradish Peroxidase Isozyme C3 Gene in Pichia pastoris[J].Biotechnology,2005,15(6):8-10. |
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| Authors: | SHAO Jin-hui ZHU You-ming HAN Jin-xiang WU Wei-ping WU Jian-mei |
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| Institution: | Shandong Medicinal Biotechnology Centre, Key Laboratory for Biotech- Drugs Ministry of Health, Jinan 250062, China |
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| Abstract: | Objective:To clone and express the horseradish peroxidase isozyme C3(HRPC3) gene.Methods: The HRPC3 gene was amplified with PCR from the genomic DNA of horseradish,and it was then linked into pMD-18T vector.After it was confirmed by sequencing analysis,it was correctly inserted into the expression vector pPIC9K of Pichia pastoris.Then the recombined pPIC9K plasmid with target gene was expressed and analyzed.Results: The HRPC3 gene was expressed successfully by using Pichia pastoris as host strain and the activity of it is 56 IU/L.Conclusion: The HRPC3 with biological activity was expressed directly in Pichia pastoris. |
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| Keywords: | horseradish peroxidase gene PCR Pichia pastoris |
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