Bonifacio G F, Brown T, Conn G L, Lane A N
Division of Molecular Structure, National Institute for Medical Research, London, England.
Biophys J. 1997 Sep;73(3):1532-8. doi: 10.1016/S0006-3495(97)78185-2.
The electrophoretic behavior of defined DNA and RNA oligonucleotide duplexes from 10 to 20 bp in length has been investigated as a function of salt conditions, gel concentration, and temperature. The RNA oligomers migrated much more slowly than the DNA oligomers of the same sequence under all conditions. From sedimentation equilibrium and velocity measurements, the apparent partial specific volume in 0.1 M KCI, 20 mM NaPi, pH 7, was determined as 0.56 +/- 0.015 ml g(-1) for DNA and 0.508 ml g(-1) for RNA. The translational friction coefficients were determined and compared with the values calculated for cylinders. Taking into account the shape factors, the solution density, and partial specific volumes, the effective degree of hydration was estimated as 0.8-1 g g(-1) DNA. There was no significant difference in the frictional coefficients of the DNA and RNA oligomers, indicating that the effective sizes of DNA and RNA are very similar in solution. The differential electrophoretic mobility of DNA and RNA must arise from the differences in interaction with counterions, which is probably a global property of the oligonucleotides.
研究了长度为10至20个碱基对的特定DNA和RNA寡核苷酸双链体的电泳行为,该行为是盐条件、凝胶浓度和温度的函数。在所有条件下,RNA寡聚物的迁移速度都比相同序列的DNA寡聚物慢得多。通过沉降平衡和速度测量,在0.1 M KCl、20 mM NaPi、pH 7条件下,DNA的表观比容测定为0.56±0.015 ml g⁻¹,RNA为0.508 ml g⁻¹。测定了平移摩擦系数,并与圆柱体计算值进行比较。考虑形状因子、溶液密度和比容,估计DNA的有效水合度为0.8 - 1 g g⁻¹。DNA和RNA寡聚物的摩擦系数没有显著差异,表明DNA和RNA在溶液中的有效尺寸非常相似。DNA和RNA的电泳迁移率差异必定源于与抗衡离子相互作用的差异,这可能是寡核苷酸的整体性质。