Käser Michael, Ruf Marie-Thérèse, Hauser Julia, Marsollier Laurent, Pluschke Gerd
Swiss Tropical Institute, Molecular Immunology, Socinstr. 57, 4002 Basel, Switzerland.
Appl Environ Microbiol. 2009 Jan;75(2):414-8. doi: 10.1128/AEM.01358-08. Epub 2008 Dec 1.
Genomic studies on pathogenic and environmental mycobacteria are of growing interest for understanding of their evolution, distribution, adaptation, and host-pathogen interaction. Since most mycobacteria are slow growers, material from in vitro cultures is usually scarce. The robust mycobacterial cell wall hinders both experimental cell lysis and efficient DNA extraction. Here, we compare elements of several DNA preparation protocols and describe a method that is economical and practical and reliably yields large amounts--usually 10-fold increased compared to earlier protocols--of highly pure genomic DNA for sophisticated downstream applications. This method was optimized for cultures of a variety of pathogenic and environmental mycobacterial species and proven to be suitable for direct mycobacterial DNA extraction from infected insect specimens.
对致病性和环境分枝杆菌进行基因组研究，对于了解它们的进化、分布、适应性以及宿主与病原体的相互作用，正变得越来越重要。由于大多数分枝杆菌生长缓慢，体外培养获得的材料通常很少。坚固的分枝杆菌细胞壁既阻碍了实验性细胞裂解，也妨碍了高效的DNA提取。在这里，我们比较了几种DNA制备方案的要素，并描述了一种经济实用的方法，该方法能可靠地产生大量——通常比早期方案增加10倍——高纯度的基因组DNA，用于复杂的下游应用。该方法针对多种致病性和环境分枝杆菌物种的培养进行了优化，并已证明适用于从受感染昆虫标本中直接提取分枝杆菌DNA。
