• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Chinese science bulletin >Effects of Li~+ and PEG on DNA uptake in yeast
【24h】

Effects of Li~+ and PEG on DNA uptake in yeast

机译：Li〜+和PEG对酵母DNA摄取的影响

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

DNA uptake of Saccharomyces cerevisiae known as genetic transformation was firstly described by Gppe-noorth in 1960, and now the most commonly used efficient protocol for yeast transformation makes use of PEG and Li~+, which works well for most laboratory strains and is suitable for high-efficiency transformation of plasmid DNA. However, it is still unknown how plasmid DNA enters yeast cells and what roles Li~+ and PEG play on DNA uptake in yeast cells until now. The elucidation of the manner in which plasmid DNA enters the intact yeast cell is beneficial for optimization of the method of yeast transformation, and more important for general understanding of the processes of macromolecule uptake by cells, especially eukaryotic cells and the mechanism of horizontal gene transfer (HGT) in eukaryotic organisms. Previous study~(151) has supposed that DNA uptake of bacteria could be divided into three steps: DNA binding to cell surfaces, penetrating through cell wall/membrane into the interior of cells, and then replicating or transcripting to RNA in the cells. To investigate the relationship between DNA binding to cell surfaces and DNA transformed into yeast cells, fluorescence microscopy was used to show the binding of pUC18 DNA labeled with the cell-impermeable YOYO-1 to cell surfaces, and the transformation frequencies of pAJ161 DNA in yeast cells were calculated. PEG was indispensable for the binding of plasmid DNA to the surfaces of intact yeast cells and protoplasts (the cell wall-removed yeast cells). Without PEG, no labeled DNA was found on the surfaces of intact yeast cells and yeast protoplasts (data not shown). The main role of PEG in the transformation is supposed to induce DNA binding to cell surfaces, and most probably to facilitate DNA to enter the cells, besides inducing membrane fusion.
机译：Gppe-noorth于1960年首次描述了酿酒酵母对DNA的吸收，即遗传转化，现在最常用的高效酵母转化方案是使用PEG和Li〜+，该方法对大多数实验室菌株都适用，适用于质粒DNA的高效转化。然而，至今仍不清楚质粒DNA如何进入酵母细胞以及Li〜+和PEG在酵母细胞DNA摄取中起什么作用。阐明质粒DNA进入完整酵母细胞的方式有助于优化酵母转化方法，并且对于一般理解细胞（尤其是真核细胞）吸收大分子的过程以及水平基因转移的机制更重要。 （HGT）在真核生物中。先前的研究[151]认为细菌对DNA的吸收可以分为三个步骤：DNA与细胞表面的结合，通过细胞壁/膜的渗透进入细胞内部，然后在细胞中复制或转录为RNA。为了研究DNA结合到细胞表面与转化到酵母细胞中的DNA之间的关系，使用荧光显微镜显示了不可渗透的YOYO-1标记的pUC18 DNA与细胞表面的结合，以及pAJ161 DNA在酵母中的转化频率计算细胞。 PEG对于质粒DNA与完整酵母细胞和原生质体（去除细胞壁的酵母细胞）表面的结合是必不可少的。没有PEG，在完整的酵母细胞和酵母原生质体的表面上没有发现标记的DNA（数据未显示）。 PEG在转化中的主要作用应该是诱导DNA与细胞表面的结合，并且最有可能促进DNA进入细胞，除了诱导膜融合。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号