Soil colloids-bound plasmid DNA: Effect on transformation of E. coli and resistance to DNase I degradation |
| |
| Authors: | P Cai W Chen K Wang W Liang |
| |
| Institution: | a State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China
b Key Laboratory of Subtropical Agricultural Resources and Environment, Huazhong Agricultural University, Wuhan 430070, China |
| |
| Abstract: | The adsorption and binding of plasmid p34S DNA on four different colloidal fractions from a Brown soil and clay minerals in the presence of various Ca2+ concentrations, the ability of bound DNA to transform competent cells of CaCl2-treated Escherichia coli, and the resistance of bound DNA to degradation by DNase I were studied. DNA adsorption on soil colloids and clay minerals was promoted in the presence of Ca2+. Kaolinite exhibited the highest adsorption affinity for DNA among the examined soil colloids and clay minerals. In comparison with organo-mineral complexes (organic clays) and fine clays (<0.2 μm), DNA was tightly adsorbed by H2O2-treated clays (inorganic clays) and coarse clays (0.2-2 μm). The transformation efficiency of bound DNA increased with increasing concentrations of Ca2+ at which soil colloid or clay mineral-DNA complexes were formed. DNA bound by kaolinite showed the lowest transformation efficiency, and especially no transformants were observed with kaolinite-DNA complex prepared at 5-100 mM Ca2+. Compared to organic clays and fine clays, DNA bound on inorganic clays and coarse clays showed a lower capacity to transform E. coli at different Ca2+ concentrations. The presence of soil colloids and minerals provided protection to DNA against degradation by DNase I. Montmorillonite, organic clays and fine clays showed stronger protective effects for DNA than inorganic clays and coarse clays. The protection mechanisms as well as the differences in transforming efficiency of plasmid DNA molecules bound on various soil colloidal particles are discussed. The information obtained in this study is of fundamental significance for the understanding of the horizontal dissemination of recombinant DNA and the fate of extracellular DNA in soil environments. |
| |
| Keywords: | Plasmid DNA Soil colloid Escherichia coli Transformation |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
