Zheng Hu-Zhi, Pang Dai-Wen, Lu Zhe-Xue, Zhang Zhi-Ling, Xie Zhi-Xiong
College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, P.R. China.
Biophys Chem. 2004 Dec 1;112(1):27-33. doi: 10.1016/j.bpc.2004.06.011.
A fluorescence microscope (FM) coupled with an intensified charge-coupled device (ICCD) camera was used to investigate the combing of DNA on cetyltrimethyl ammonium bromide (CTAB)-coated glass surfaces. DNA molecules can be combed uniform and straight on CTAB-coated surfaces. Different combing characteristics at different pH values were found. At lower pH (ca. 5.5), DNA molecules were stretched 30% longer than the unextended and DNA extremities bound with CTAB-coated surfaces via hydrophobic interaction. At high pH values (e.g., 6.4 and 6.5), DNA molecules were extended about 10% longer and DNA extremities bound with CTAB-coated surfaces via electrostatic attraction. At pH 6.0, DNA molecules could be extended 30% longer on 0.2-mM CTAB-coated surfaces. CTAB cationic surfactant has both a hydrophobic motif and a positively charged group. So, CTAB-coated surfaces can bind DNA extremities via hydrophobic effect or electrostatic attraction at different pH values. It was also found that combing of DNA on CTAB-coated surfaces is reversible. The number of DNA base pairs binding to CTAB-coated surfaces was calculated.
使用一台与增强型电荷耦合器件(ICCD)相机联用的荧光显微镜(FM)来研究十六烷基三甲基溴化铵(CTAB)包被的玻璃表面上DNA的梳理情况。DNA分子能够在CTAB包被的表面上被均匀且笔直地梳理。发现了在不同pH值下的不同梳理特性。在较低pH(约5.5)时,DNA分子比未伸展时长30%,并且DNA末端通过疏水相互作用与CTAB包被的表面结合。在高pH值(例如6.4和6.5)时,DNA分子伸展约10%,并且DNA末端通过静电吸引与CTAB包被的表面结合。在pH 6.0时,DNA分子在0.2 mM CTAB包被的表面上能够伸展30%。CTAB阳离子表面活性剂既有疏水基团又有带正电荷的基团。所以,CTAB包被的表面能够在不同pH值下通过疏水作用或静电吸引结合DNA末端。还发现DNA在CTAB包被的表面上的梳理是可逆的。计算了与CTAB包被的表面结合的DNA碱基对数量。