Amplification of plasmid DNA bound on soil colloidal particles and clay minerals by the polymerase chain reaction  

作　　者：CAI Peng HUANG Qiao-yun LU Yan-du CHEN Wen-li JIANG Dai-hua LIANG Wei 

机构地区：[1]State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China [2]Key Laboratory of Subtropical Agricultural Resources a~ld Environment, Huazhong Agricultural University, Wuhan 430070, China

出　　处：《Journal of Environmental Sciences》2007年第11期1326-1329,共4页环境科学学报（英文版）

基　　金：Project supported by the National Natural Science Foundation of China(No.40271064)

摘　　要：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.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.

关 键 词：ADSORPTION AMPLIFICATION MINERAL PCR plasmid DNA soil colloid 

分 类 号：TQ424[化学工程]