Liao Yu-bo, Meng Yao-yong, Lei Hao-dong, Wang Ying
Photonic Chinese Medicine Lab, South China Normal University, Guangzhou 510631, China.
Guang Pu Xue Yu Guang Pu Fen Xi. 2007 May;27(5):940-3.
The Raman spectra of PolyA, PolyU and their double-stranded complex were measured, and the spectral changes upon the formation of double-stranded complex were studied. The experimental results show: (1) Under the experimental conditions used in the present work (0.14 mol x L(-1) NaCl, 1 mmol x L(-1) Tris solution, neutral pH and 15 degrees C), PolyU, PolyA and PolyA x PolyU occur as random-coiled, A-single-stranded helical and A-double-stranded helical conformations, respectively. One of the main spectral differences between the latter two conformations and the former one is the Raman band near 814 cm(-1) of ordered structures. Another difference is in the full width at half the maximum (i.e. FWHM) of the band near 1100 cm(-1). The FWHM of the band 1100 cm(-1) of PolyA is the same as that of PolyA x PolyU, while the band of PolyU shows remarkable broadening. In addition, we found that the conformation of PolyA is somewhat not so ordered as that of its duplex, which can be concluded from the value of I814/I1100 of the two polynucleotides. (2) The formation of duplex makes base-base stacking interactions much stronger, and the conformation of the backbone more ordered, which leads to obvious Raman hypochromic effect with some corresponding band shift. In this process, PolyU underwent more significant spectral changes than PolyA. As spectral markers, these results can be of great importance in Raman spectral signal detection of gene-chips.
测量了聚腺苷酸(PolyA)、聚尿苷酸(PolyU)及其双链复合物的拉曼光谱,并研究了双链复合物形成时的光谱变化。实验结果表明:(1)在本工作所采用的实验条件下(0.14 mol·L⁻¹氯化钠、1 mmol·L⁻¹ Tris溶液、中性pH值和15℃),PolyU、PolyA和PolyA×PolyU分别以无规卷曲、A单链螺旋和A双链螺旋构象存在。后两种构象与前一种构象之间的主要光谱差异之一是有序结构在814 cm⁻¹附近的拉曼谱带。另一个差异在于1100 cm⁻¹附近谱带的半高宽(即FWHM)。PolyA的1100 cm⁻¹谱带的FWHM与PolyA×PolyU的相同,而PolyU的谱带则显著变宽。此外,我们发现从两种多核苷酸的I814/I1100值可以得出,PolyA的构象不如其双链体的构象有序。(2)双链体的形成使碱基 - 碱基堆积相互作用更强,主链构象更有序,这导致明显的拉曼减色效应并伴有一些相应的谱带位移。在此过程中,PolyU比PolyA经历了更显著的光谱变化。作为光谱标记,这些结果在基因芯片的拉曼光谱信号检测中可能具有重要意义。