Zainuddin Z F, Kunze Z M, Dale J W
Department of Microbiology, University of Surrey, Guildfor, UK.
Mol Microbiol. 1989 Jan;3(1):29-34. doi: 10.1111/j.1365-2958.1989.tb00100.x.
One limiting factor in studies of tuberculosis and leprosy is the difficulty of genetic analysis and manipulation of mycobacteria. Two approaches were adopted for the construction of vectors, based on different Escherichia coli plasmids and using Mycobacterium smegmatis as the host. In both cases we found that the original E. coli plasmid is capable of being replicated in M. smegmatis, yielding chloramphenicol-resistant colonies. One such plasmid has been recovered from a M. smegmatis transformant and used to re-transform both M. smegmatis and E. coli to chloramphenicol resistance. This plasmid is indistinguishable from the original plasmid by restriction analysis, and can be used as a shuttle vector for the genetic manipulation of mycobacterial species.
结核病和麻风病研究中的一个限制因素是分枝杆菌基因分析和操作的困难。基于不同的大肠杆菌质粒并以耻垢分枝杆菌作为宿主，采用了两种方法构建载体。在这两种情况下，我们都发现原始的大肠杆菌质粒能够在耻垢分枝杆菌中复制，产生抗氯霉素菌落。已从耻垢分枝杆菌转化体中回收了一种这样的质粒，并用于将耻垢分枝杆菌和大肠杆菌再次转化为抗氯霉素。通过限制性分析，该质粒与原始质粒无法区分，可作为用于分枝杆菌物种基因操作的穿梭载体。
