重金属复合污染农田土壤DNA的快速提取及其PCR-DGGE分析
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* 国家杰出青年基金项目(40125005);国家重点基础研究发展规划项目(2002CB410810、2002CB410809);中国科学院知识创新工程重要方向项目(KZCX3-SW-429)资助


RAPID EXTRACTION AND PURIFICATION OF DNA IN FARMLAND SOILS CONTAMINATED WITH MIXED HEAVY METALS FOR PCR-DGGE ANALYSIS
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    摘要:

    国内首次运用FastPrep®核酸快速提取系统提取了重金属复合污染农田土壤的DNA,并对其进行了聚合酶链反应—变性梯度凝胶电泳(PCR-DGGE)分析。结果表明,FastPrep®核酸提取仪与相应的FastDNA SPIN Kit for Soil试剂盒联用时,能有效地分离到纯度较高的重金属污染农田土壤的DNA。PCR-DGGE电泳图谱表明,PCR产物经DGGE检测后得到的电泳条带清晰且分离效果好,可以明显反映出重金属复合污染导致了农田土壤微生物在基因上的损伤,影响到农田土壤生态系统的细菌丰富度,改变了土壤环境的优势菌群,从而使农田土壤微生物群落结构多样性发生变化。可见,FastPrep®核酸提取系统同样适用于重金属污染农田土壤环境中微生物基因组DNA的快速分离和纯化,得到的DNA可直接用于PCR-DGGE分析。

    Abstract:

    Total community DNA was extracted and purified from soils contaminated with different levels of mixed heavy metals by FastPrep ®system, and then amplified by using eubacterial 16S rDNAs primers.Their PCR products were analysed by DGGE to obtain bacterial community patterns.The results showed that the FastPrep ®120 instrument with the FastDNA SPIN Kit for soil can be used to rapidly, effectively extract and purify DNA from polluted soils.The electrophoresis lanes obtained by PCR-DGGE were legible and distinguishable.The profiles of PCR-DGGE indicated that heavy metals accumulat ion at different levels markedly caused damage to farmland soils DNA, affecting the soil microbial richness and structure of dominating bacterial populations in the farmland ecological systems, and changing the structural diversity of microbial community.The FastPrep ®system is also suitable for rapid efficient extraction and purification of soil total community DNA from farmland soils polluted with mixed heavy metals and in turn the DNA can be used directly for PCR-DGGE analysis.

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滕应,骆永明,赵祥伟,李振高,宋静,吴龙华.重金属复合污染农田土壤DNA的快速提取及其PCR-DGGE分析[J].土壤学报,2004,41(3):343-347. DOI:10.11766/trxb200312230303 Teng Ying, Luo Yongming, Zhao Xiangwei, Li Zhengao, Song Jing, Wu Longhua. RAPID EXTRACTION AND PURIFICATION OF DNA IN FARMLAND SOILS CONTAMINATED WITH MIXED HEAVY METALS FOR PCR-DGGE ANALYSIS[J]. Acta Pedologica Sinica,2004,41(3):343-347.

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  • 收稿日期:2003-12-23
  • 最后修改日期:2004-01-24
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  • 在线发布日期: 2013-02-25
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