Zheng Hu-Zhi, Liu Hui-Hui, Chen Shao-Xing, Lu Zhe-Xue, Zhang Zhi-Ling, Pang Dai-Wen, Xie Zhi-Xiong, Shen Ping
College of Chemistry & Molecular Sciences and College of Life Sciences, Wuhan University, Wuhan 430072, P. R. China.
Bioconjug Chem. 2005 Mar-Apr;16(2):250-4. doi: 10.1021/bc049833v.
A new method based on fluorescence imaging and flow cytometry was developed to investigate the transformation process of Saccharomyces cerevisiae AY. Yeast and fluorescent-labeled plasmid pUC18 were used as models of cells and DNA molecules, respectively. Binding of DNA molecules to yeast cell surfaces was observed. Factors influencing DNA binding to cell surfaces were investigated. It has been found that poly(ethylene glycol) (PEG) could induce DNA binding to yeast surfaces, while Li(+) showed a weak effect on the binding. When both Li(+) and PEG were used, synergetic effect occurred, resulting in the binding of pUC18 to the surface of more yeast cells compared with that in the presence of PEG or Li(+) only. It was also confirmed that heat shock, Li(+), and PEG all can increase the permeability of yeast cells. This simple method is helpful for understanding the process of yeast transformation and can be used to investigate the interaction of DNA with cell surfaces.
开发了一种基于荧光成像和流式细胞术的新方法来研究酿酒酵母AY的转化过程。酵母和荧光标记的质粒pUC18分别用作细胞和DNA分子的模型。观察到DNA分子与酵母细胞表面的结合。研究了影响DNA与细胞表面结合的因素。已发现聚乙二醇（PEG）可诱导DNA与酵母表面结合，而Li(+)对结合的影响较弱。当同时使用Li(+)和PEG时，会产生协同效应，导致与仅存在PEG或Li(+)时相比，更多酵母细胞表面结合有pUC18。还证实热休克、Li(+)和PEG均可增加酵母细胞的通透性。这种简单的方法有助于理解酵母转化过程，并可用于研究DNA与细胞表面的相互作用。
