一种快速提取细菌总DNA的方法研究

随着分子生物学技术应用于环境微生物研究的深入开展,占自然界微生物物种总数的90%以上的不能人工培养或培养困难的微生物已经可以借助分子生物学技术进行功能基因的开发和利用。而快速得到纯度较高,结构完整的细菌染色体DNA成为这一技术得以实现的前提。本文报道了利用高温处理和SDS的裂解作用相结合而建立的一种快速、简便的提取细菌染色体DNA的方法。经过脉冲电泳实验证明,利用本方法提取得到的几种革兰氏阳性和革兰氏阴性菌株的基因组DNA结构完整,并且无明显降解,无须经过纯化,可以直接进行PCR扩增和酶切等分子生物学操作,将此方法进一步应用于土壤环境DNA的提取方面,同样达到了快速得到大片段、高质量的环境微生物基因组的目的,为研究未培养的环境微生物多样性打下了坚实的基础,同时为环境基因组的提取提供了一个新的途径。

A speedy extraction method for bacterial chromosomal DNA

How to get functional gene from uncultured-microbiology is the hotspot content of microbial ecology. What the most important is how to obtain the pure and integrated genomic DNA. This article had showed an efficient, nonselective extraction method to gain chromosomal DNA from eight kinds of bacteria. Amount DNA released by hot-detergent gave the highest DNA yields from different G+ and G- bacteria. Running 20 hours by PFGE mode, the size of total DNA is over 23kb. The pure DNA could be digested by Hind III and used in PCR. The total environmental DNA also can be extracted from soil by the same method. As a result it showed a new way for the environmental DNA extraction.