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模型B-DNA寡聚物d(CGCGAATTCGCG)2和活鲑鱼精子中的DNA的拉曼光谱表明,两者都具有非常相似的B型构象。

Raman spectra of the model B-DNA oligomer d(CGCGAATTCGCG)2 and of the DNA in living salmon sperm show that both have very similar B-type conformations.

作者信息

Kubasek W L, Wang Y, Thomas G A, Patapoff T W, Schoenwaelder K H, Van der Sande J H, Peticolas W L

出版信息

Biochemistry. 1986 Nov 18;25(23):7440-5. doi: 10.1021/bi00371a028.

Abstract

Raman spectra were obtained from aqueous solutions of the deoxyoligonucleotide d(CGCGAATTCGCG)2 (I), which has been suggested as a model for B-type DNA conformation. These spectra were compared with the Raman spectra of the aqueous solutions of several DNAs of natural origin taken under identical solution conditions. Since the model sequence has a high percent GC (66%), the Raman spectrum was compared with the Raman spectrum of the DNA from Micrococcus lysodeikticus (72% GC), and the spectra of the two different DNAs were found to be rather similar in both 50 mM salt and 6 M salt solutions. Computer-aided band-shape analysis of the backbone vibrational region of the Raman spectra shows the existence of several bands corresponding to different furanose ring puckers. This appears to indicate a heterogeneity of furanose ring pucker in both the model dodecamer and the native DNA. Significant differences were found in the intensity of the conformational marker band at 810 cm-1, which indicates corresponding differences in furanose ring pucker heterogeneities in these two high GC content DNAs. The Raman spectrum of the dodecamer (I) was used to analyze the Raman spectrum of the DNA inside the head of living intact salmon sperm. Sperm spectra were taken with both our conventional Raman spectrograph and a newly developed intracavity laser Raman microscope system. Although the DNA in the sperm head is required by packing considerations to be in a highly compact and condensed state, the Raman spectra of the intact sperm are almost identical with that of the model dodecamer (I) if the difference in base composition is taken into account.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

拉曼光谱是从脱氧寡核苷酸d(CGCGAATTCGCG)2 (I)的水溶液中获得的,该脱氧寡核苷酸被认为是B型DNA构象的模型。这些光谱与在相同溶液条件下获取的几种天然来源DNA水溶液的拉曼光谱进行了比较。由于模型序列具有较高的GC百分比(66%),因此将拉曼光谱与来自溶壁微球菌的DNA的拉曼光谱(GC含量为72%)进行了比较,发现在50 mM盐溶液和6 M盐溶液中,这两种不同DNA的光谱相当相似。对拉曼光谱主链振动区域的计算机辅助带形分析表明,存在几个对应于不同呋喃糖环褶皱的谱带。这似乎表明在模型十二聚体和天然DNA中呋喃糖环褶皱存在异质性。在810 cm-1处的构象标记带强度上发现了显著差异,这表明这两种高GC含量DNA中呋喃糖环褶皱异质性存在相应差异。十二聚体(I)的拉曼光谱被用于分析完整活鲑鱼精子头部内DNA的拉曼光谱。精子光谱是用我们传统的拉曼光谱仪和新开发的腔内激光拉曼显微镜系统采集的。尽管从堆积角度考虑,精子头部的DNA需要处于高度紧凑和浓缩的状态,但如果考虑碱基组成的差异,完整精子的拉曼光谱与模型十二聚体(I)的几乎相同。(摘要截短于250字)

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