Gray D M, Edmondson S P, Lang D, Vaughan M
Nucleic Acids Res. 1979;6(6):2089-107. doi: 10.1093/nar/6.6.2089.
It is known that DNA in aqueous-ethanol solutions undergoes a B to A conformational change between 60% and 80% (w/w) ethanol. We have found that precipitates formed by adding salt to DNA in 60% and 80% ethanolic solutions can be very different. DNA precipitated from 60% ethanol forms a fine condensate that only slowly settles out of suspension and shows a characteristic differential scattering of circularly polarized light at long wavelengths. DNA precipitated from 80% ethanol forms a flocculent aggregate that exhibits the CD spectral features of the A conformation. Data from circular dichroism spectra of natural and synthetic nucleic acids and from X-ray diffraction patterns of the precipitates show that DNA molecules precipitated from 60% and 80% ethanol are, respectively, in the B and A conformation. Therefore, the different secondary conformations of DNA in ethanolic solutions are maintained during precipitation under these conditions. These results are of general importance for the preparation and study of condensed forms of DNA, since a relatively small change in the extent of dehydration can change the secondary conformation of DNA and markedly affect the character of a subsequent precipitate.
已知在乙醇水溶液中,当乙醇含量在60%至80%(w/w)之间时,DNA会发生从B构象到A构象的转变。我们发现,向60%和80%乙醇溶液中的DNA添加盐所形成的沉淀物可能会有很大差异。从60%乙醇中沉淀出的DNA形成一种精细的凝聚物,它仅缓慢地从悬浮液中沉降出来,并且在长波长处呈现出圆偏振光的特征性差分散射。从80%乙醇中沉淀出的DNA形成一种絮状聚集体,它展现出A构象的圆二色光谱特征。天然和合成核酸的圆二色光谱数据以及沉淀物的X射线衍射图谱表明,从60%和80%乙醇中沉淀出的DNA分子分别处于B构象和A构象。因此,在这些条件下沉淀过程中,乙醇溶液中DNA的不同二级构象得以保持。这些结果对于DNA凝聚形式的制备和研究具有普遍重要性,因为脱水程度相对较小的变化就可以改变DNA的二级构象,并显著影响后续沉淀物的特性。