Amplification of plasmid DNA bound on soil colloidal particles and clay
minerals by the polymerase chain reaction |
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Authors: | CAI Peng HUANG Qiao-yun LU Yan-du CHEN Wen-li JIANG Dai-hua and LIANG Wei |
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Institution: | 1. State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China 2. State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China;Key Laboratory of Subtropical Agricultural Resources and Environment, Huazhong Agricultural University, Wuhan 430070, China |
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Abstract: | Polymerase chain reaction (PCR) was used to amplify a 600-base pair (bp) sequence of plasmid pGEX-2T DNA bound on soil colloidal particles from Brown soil (Alfisol) and Red soil (Ultisol), and three different minerals (goethite, kaolinite, montmorillonite). DNA bound on soil colloids, kaolinite, and montmorillonite was not amplified when the complexes were used directly but amplification occurred when the soil colloid or kaolinite-DNA complex was diluted, 10- and 20-fold. The montmorillonite-DNA complex required at least 100-fold dilution before amplification could be detected. DNA bound on goethite was amplified irrespective of whether the complex was used directly, or diluted 10- and 20-fold. The amplification of mineral-bound plasmid DNA by PCR is, therefore, markedly influenced by the type and concentration of minerals used. This information is of fundamental importance to soil molecular microbial ecology with particular reference to monitoring the fate of genetically engineered microorganisms and their recombinant DNA in soil environments. |
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Keywords: | adsorption amplification mineral PCR plasmid DNA soil colloid polymerase chain reaction clay minerals colloidal particles soil colloids bound plasmid DNA information fundamental importance molecular ecology reference monitoring fate microorganisms recombinant environments type concentration dilution |
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