| 质粒DNA的阴离子交换色谱法纯化及内毒素去除 |
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| 引用本文： | 皮文辉,孙从建,宋志强,马连营,刘淑萍,白丁平,蔡宜东,刘守仁.质粒DNA的阴离子交换色谱法纯化及内毒素去除[J].色谱,2007,25(6):809-813. |
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| 作者姓名： | 皮文辉 孙从建 宋志强 马连营 刘淑萍 白丁平 蔡宜东 刘守仁 |
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| 作者单位： | 1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China; 2.Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China; 3.Xinjiang Tianwu Science & Technology Development Co., Ltd., Urumqi 830011, China |
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| 摘 要： | 采用Fractogel EMD TMAE(M)强阴离子交换介质分离纯化质粒脱氧核糖核酸（DNA），该介质对质粒DNA的动态载样量达0.62 mg/mL。用Triton X-114或Triton X-100预先处理质粒DNA的裂解澄清液后，经阴离子交换色谱分离纯化获得的质粒DNA中内毒素含量分别为6.42 EU/mg和9.50 EU/mg,显著低于未经Triton处理的裂解澄清液（67.82 EU/mg）。该法实现了阴离子交换色谱一步纯化质粒DNA的目的,具有简便、省时、成本低等特点。
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| 关 键 词： | 阴离子交换色谱 纯化 质粒DNA Triton X-114 Triton X-100 内毒素 去除 |
| 文章编号： | 1000-8713（2007）06-0809-05 |
| 收稿时间： | 21 May 2007 |
| 修稿时间： | 2007-05-21 |
Purification of Plasmid DNA Using Anion-Exchange Chromatography and Removal of Endotoxin |
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| PI Wenhui,SUN Congjian,SONG Zhiqiang,MA Lianying,LIU Shuping,BAI Dingping,CAI Yidong,LIU Shouren.Purification of Plasmid DNA Using Anion-Exchange Chromatography and Removal of Endotoxin[J].Chinese Journal of Chromatography,2007,25(6):809-813. |
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| Authors: | PI Wenhui SUN Congjian SONG Zhiqiang MA Lianying LIU Shuping BAI Dingping CAI Yidong LIU Shouren |
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| Institution: | 1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China; 2.Institute of Animal Husbandry and Veterinary Medicine, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China; 3.Xinjiang Tianwu Science & Technology Development Co., Ltd., Urumqi 830011, China |
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| Abstract: | The plasmid DNA was purified using the Fractogel EMD TMAE (M) anion exchange media. The dynamic loading capacity of this media for plasmid DNA was 0.62 mg/mL. It was found that when the lysate of bacteria was incubated with either Triton X-114 or Triton X-100 before purification, the average content of endotoxin left in the purified plasmid DNA was 6.42 EU/mg or 9.50 EU/mg, respectively. These results were much lower than that without pre-incubation (67.82 EU/mg). The purification of plasmid DNA and the removal of endotoxin were achieved in one step using anion-exchange chromatography. This method is simple, fast and of low cost. |
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| Keywords: | Triton X-114 Triton X-100 |
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