We investigated the interaction between DNA and hexammine cobalt III [Co（NH3）6]3＋ by a simple molecular combing method and dynamic light scattering. The average extension of A- DNA-YOYO-1 complex is found to be 20....We investigated the interaction between DNA and hexammine cobalt III [Co（NH3）6]3＋ by a simple molecular combing method and dynamic light scattering. The average extension of A- DNA-YOYO-1 complex is found to be 20.9μm, about 30% longer than the contour length of the DNA in TE buffer （10 mmol/L Tris, 1 mmol/L EDTA, pH=8.0）, due to bis-intercalation of YOYO-1. A multivalent cation, hexammine cobalt, is used for DNA condensation. We find that the length of DNA-[Co（NH3）6]3＋ complexes decrease from 20.9 μm to 5.9μm as the concentration of the [Co（NH3）6]3＋ vary from 0 to 3 μmol/L. This observation provides a direct visualization of single DNA condensation induced by hexammine cobalt. The results from the molecular combing studies are supported by dynamic light scattering investigation, where the average hydrodynamic radius of the DNA complex decreases from 203.8 nm to 39.26 nm under the same conditions. It shows that the molecular combing method is feasible for quantitative conformation characterization of single bio-macromolecules.展开更多
文摘We investigated the interaction between DNA and hexammine cobalt III [Co（NH3）6]3＋ by a simple molecular combing method and dynamic light scattering. The average extension of A- DNA-YOYO-1 complex is found to be 20.9μm, about 30% longer than the contour length of the DNA in TE buffer （10 mmol/L Tris, 1 mmol/L EDTA, pH=8.0）, due to bis-intercalation of YOYO-1. A multivalent cation, hexammine cobalt, is used for DNA condensation. We find that the length of DNA-[Co（NH3）6]3＋ complexes decrease from 20.9 μm to 5.9μm as the concentration of the [Co（NH3）6]3＋ vary from 0 to 3 μmol/L. This observation provides a direct visualization of single DNA condensation induced by hexammine cobalt. The results from the molecular combing studies are supported by dynamic light scattering investigation, where the average hydrodynamic radius of the DNA complex decreases from 203.8 nm to 39.26 nm under the same conditions. It shows that the molecular combing method is feasible for quantitative conformation characterization of single bio-macromolecules.
