Comprehensive Identification of Salmonella enterica Serovar Typhimurium Genes Required for Infection of BALB/c Mice

PLoS Pathogens

Paul J. Owen¹  Andrew G. Allen¹  Gil Shalom¹  Dov J. Stekel²  Ian G. Charles³  Chrissie Willers⁴  Stephen J. Pleasance⁴  Sarah E. Peters⁴  Gavin K. Paterson⁴  Danielle B. Cone⁴  Duncan J. Maskell⁴  Roy R. Chaudhuri⁴  Helen Northen⁴ 

[1] Arrow Therapeutics Ltd., London, United Kingdom;Centre for Systems Biology, School of Biosciences, University of Birmingham, Edgbaston, Birmingham, United Kingdom;Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, United Kingdom;Department of Veterinary Medicine, University of Cambridge, Cambridge, United Kingdom

关键词: Transposable elements;    Microarrays;    Salmonella typhimurium;    Gene pool;    Mammalian genomics;    Mouse models;    Deletion mutagenesis;    Salmonellosis;   

DOI  :  10.1371/journal.ppat.1000529

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Genes required for infection of mice by Salmonella Typhimurium can be identified by the interrogation of random transposon mutant libraries for mutants that cannot survive in vivo. Inactivation of such genes produces attenuated S. Typhimurium strains that have potential for use as live attenuated vaccines. A quantitative screen, Transposon Mediated Differential Hybridisation (TMDH), has been developed that identifies those members of a large library of transposon mutants that are attenuated. TMDH employs custom transposons with outward-facing T7 and SP6 promoters. Fluorescently-labelled transcripts from the promoters are hybridised to whole-genome tiling microarrays, to allow the position of the transposon insertions to be determined. Comparison of microarray data from the mutant library grown in vitro (input) with equivalent data produced after passage of the library through mice (output) enables an attenuation score to be determined for each transposon mutant. These scores are significantly correlated with bacterial counts obtained during infection of mice using mutants with individual defined deletions of the same genes. Defined deletion mutants of several novel targets identified in the TMDH screen are effective live vaccines.

【 授权许可】

CC BY