Isolation of RNA from mycobacteria grown under in vitro and in vivo conditions
Isolation of RNA from mycobacteria grown under in vitro and in vivo conditions
复制标题
DOI：
10.1111/j.1574-6968.2000.tb09100.x
复制
发表时间：
2000-05-15
影响因子：
2.1
通讯作者：
Knapp, B
中科院分区：
文献类型：
--
作者：
Dietrich, G;Schaible, UE;Knapp, B
从分枝杆菌中分离RNA非常困难，并且产量通常很长。我们描述了一种从分枝杆菌物种中分离RNA的方法，该方法结合了在往复振荡器中通过二氧化硅/陶瓷基质对分枝杆菌细胞的破坏，以及随后在具有基于硅胶的膜的旋转柱上进行RNA纯化的简便性和效率。该方法快速，易于操作，并产生大量的纯的，完整的总RNA。由于其安全性，该方法甚至适用于第3组生物危害微生物，如结核分枝杆菌。通过将从感染的原代巨噬细胞中分离吞噬体细菌的方法与新的RNA分离技术相结合，我们能够监测感染过程中的基因表达，即使是在对通用操作具有相当抗性的细菌中，如牛分枝杆菌。(C)2000年，欧洲微生物学会联合会。由Elsevier Science B. V.出版，版权所有。
Isolation of RNA from mycobacteria is very difficult to perform, and the yields are generally very lon. We describe an approach to isolate RNA from mycobacterial species which combines the disruption of mycobacterial cells by a silica/ceramic matrix in a reciprocal shaker with the ease and efficiency of subsequent RNA purification on spin columns with silica gel-based membranes. This method is rapid, easy to perform and yields high amounts of pure, intact total RNA. Due to its safety, this method is applicable even to group 3 biological hazard organisms like Mycobacterium tuberculosis. By combining a method for the isolation of phagosomal bacteria from infected primary macrophages with the novel RNA isolation technique, we are able to monitor gene expression during infection even in bacteria which are rather resistant to generic manipulation, like Mycobacterium bovis. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.