基于矿石纳米材料DNA转化机理初探及方法改进.pdfVIP

生 物 工 程 学 报 Chin J Biotech 2010, October 25; 26(10): 1379−1384 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn ©2010 CJB, All rights reserved. 细胞工厂的构建技术 基于矿石纳米材料的DNA 转化的机理初探及方法改进 谭海东，王磊，林金涛，赵宗保 中国科学院大连化学物理研究所生物技术部，大连 116023 摘 要: 一种价格便宜、资源丰富和对人体无害的矿石纳米材料——海泡石，用于微生物DNA 转化。该法简便、快速、 对人体健康无害，但对这种转化方法机制的理解还有很多问题。通过小片段RNA 竞争试验，发现了与先前报道不同的 转化机制。同时，对该法进行了优化，结果可以实现对冷藏 1 个月的大肠杆菌Escherichia coli DH5α单菌落直接转化， 无需感受态制备和处理后的温育过程，可得到比钙转高的转化率。由于可优化的参数很多，所以这种转化方法可以提 升的空间很大。同时，该方法可用于探索其他用钙转和电转未成功的微生物。 关键词: 矿石纳米纤维材料，海泡石，DNA 转化，转化机制 Mechanism of DNA transformation based on mineral nanofibers and method improvement Haidong Tan, Lei Wang, Jintao Lin, and Zongbao (Kent) Zhao Biotechnology Division, Dalian Institute of Chemical Physics , Chinese Academy of Sciences, Dalian 116023, China Abstract: Sepiolite−an inexpensive, resourceful, fibrous yet inoffensive mineral−made DNA transformation rapid, simple and efficient but the mechanism for DNA transformation was still unclear. Through RNA competition test, we proposed the different transforming mechanisms from the previous report. Meanwhile, we optimized the transforming method and could transfer a colony stored at 4°C for a month with plasmid through sepiolite fibers. The cells could be transformed well without competent cells preparation or incubation process. In sum, this was a novel potential transforming method, which could be explored further if the chemical method and electroporation could not be used. Keywords: mineral nanofibers, sepiolite, DNA transformation, mechanism for transforming 微生物的 DNA 转化方法是基因工程中的重要内 系列关于使用温石