Hensel M, Shea J E, Gleeson C, Jones M D, Dalton E, Holden D W
Department of Infectious Diseases and Bacteriology, Royal Postgraduate Medical School, Hammersmith Hospital, London, UK.
Science. 1995 Jul 21;269(5222):400-3. doi: 10.1126/science.7618105.
An insertional mutagenesis system that uses transposons carrying unique DNA sequence tags was developed for the isolation of bacterial virulence genes. The tags from a mixed population of bacterial mutants representing the inoculum and bacteria recovered from infected hosts were detected by amplification, radiolabeling, and hybridization analysis. When applied to a murine model of typhoid fever caused by Salmonella typhimurium, mutants with attenuated virulence were revealed by use of tags that were present in the inoculum but not in bacteria recovered from infected mice. This approach resulted in the identification of new virulence genes, some of which are related to, but functionally distinct from, the inv/spa family of S. typhimurium.
为了分离细菌毒力基因，开发了一种插入诱变系统，该系统使用携带独特DNA序列标签的转座子。通过扩增、放射性标记和杂交分析来检测来自代表接种物的细菌突变体混合群体以及从感染宿主中回收的细菌的标签。当应用于鼠伤寒沙门氏菌引起的伤寒热小鼠模型时，通过使用接种物中存在但从感染小鼠中回收的细菌中不存在的标签，揭示了毒力减弱的突变体。这种方法导致了新毒力基因的鉴定，其中一些与鼠伤寒沙门氏菌的inv/spa家族相关，但在功能上有所不同。
