Amplification of plasmid DNA bound on soil colloidal particles and clay minerals by the polymerase chain reaction.
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.
Cai
Peng
P
State Key Laboratory ofAgricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China. cpcp@webmail.hzau.edu.cn
Huang
Qiao-yun
QY
Lu
Yan-du
YD
Chen
Wen-li
WL
Jiang
Dai-hua
DH
Liang
Wei
W
D000538:Aluminum Silicates / Q000737:chemistry*; D000818:Animals; D001546:Bentonite / Q000737:chemistry; D002417:Cattle; D000077215:Clay; D003102:Colloids / Q000737:chemistry*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D058085:Iron Compounds / Q000737:chemistry; D007616:Kaolin / Q000737:chemistry; D008903:Minerals; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012987:Soil
D000538:Aluminum Silicates; D003102:Colloids; D004269:DNA, Bacterial; D058085:Iron Compounds; D008903:Minerals; D012987:Soil; D001546:Bentonite; C094886:goethite; D007616:Kaolin; D000077215:Clay