Institute for Biotechnology, Bay Zoltán Foundation for Applied Research, Szeged, Hungary.
J Appl Microbiol. 2010 Nov;109(5):1845-52. doi: 10.1111/j.1365-2672.2010.04818.x. Epub 2010 Jul 30.
To develop an applicable vector system and a transformation method for the manipulation of Dietzia spp.
The pNV18 Nocardia-E. coli shuttle vector was tested and found to be a replicating plasmid in Dietzia sp. E1. With the use of pNV18, an electroporation method was optimized for the transformation of Dietzia sp. E1, and a transformation efficiency suitable for genetic manipulations was achieved (2·18×10(4) transformants μg(-1) DNA). The method was also applied for the transformation of Dietzia cinnamea, D. maris, D. natronolimnaea and D. psychralcaliphila.
The first applicable vectors and a simple electroporation protocol enabling the manipulation of several Dietzia spp. are presented.
Dietzia spp. have clinical, industrial and great environmental importance; however, the analysis of the Dietzia genus is currently hampered by the lack of manipulation techniques. The presented basic tools allow the genetic analysis of several Dietzia species, including the human disease-associated Dietzia maris.
开发适用于狄氏菌属操作的载体系统和转化方法。
检测到诺卡氏菌-大肠杆菌穿梭载体 pNV18 在狄氏菌属 E1 中可复制。利用 pNV18,优化了狄氏菌属 E1 的电穿孔转化方法,获得了适合遗传操作的转化效率(2·18×10(4)转化体 μg(-1)DNA)。该方法还适用于变色狄氏菌、海生狄氏菌、嗜碱狄氏菌和嗜冷狄氏菌的转化。
本文提出了第一个适用的载体和一种简单的电穿孔方案,可用于几种狄氏菌属的操作。
狄氏菌属具有临床、工业和重要的环境意义;然而,目前缺乏操作技术限制了对狄氏菌属的分析。本文提出的基本工具允许对包括与人类疾病相关的狄氏菌属海生狄氏菌在内的几种狄氏菌属进行遗传分析。