色谱

色谱

• 研究论文 • 上一篇    下一篇

质粒DNA的阴离子交换色谱法纯化及内毒素去除

皮文辉1,2,孙从建3,宋志强2,马连营3,刘淑萍3,白丁平2,蔡宜东3,刘守仁2

  

  1. 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
  • 收稿日期:2007-05-21 修回日期:2007-07-18 出版日期:2007-11-30 发布日期:1984-12-25
  • 通讯作者: 刘守仁

Purification of Plasmid DNA Using Anion-Exchange Chromatography and Removal of Endotoxin

PI Wenhui1,2, SUN Congjian3, SONG Zhiqiang2, MA Lianying3, LIU Shuping3, BAI Dingping2, CAI Yidong3, LIU Shouren2   

  1. 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
  • Received:2007-05-21 Revised:2007-07-18 Online:2007-11-30 Published:1984-12-25

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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。

关键词: Triton X-100, Triton X-114, 纯化, 内毒素, 去除 , 阴离子交换色谱, 质粒DNA

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.

Key words: Triton X-114, endotoxin, plasmid DNA, purification, removal , Triton X-100, anion-exchange chromatography