Defining the ABC of gene essentiality in streptococci

Charbonneau, Amelia R. L. and Forman, Oliver P. and Cain, Amy K. and Newland, Graham and Robinson, Carl and Boursnell, Mike and Parkhill, Julian and Leigh, James A. and Maskell, Duncan J. and Waller, Andrew S. (2017) Defining the ABC of gene essentiality in streptococci. BMC Genomics, 18 (1). 426/1-426/11. ISSN 1471-2164

[img]
Preview
PDF - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
Available under Licence Creative Commons Attribution.
Download (1MB) | Preview

Abstract

Background: Utilising next generation sequencing to interrogate saturated bacterial mutant libraries provides unprecedented information for the assignment of genome-wide gene essentiality. Exposure of saturated mutant libraries to specific conditions and subsequent sequencing can be exploited to uncover gene essentiality relevant to the condition. Here we present a barcoded transposon directed insertion-site sequencing (TraDIS) system to define an essential gene list for Streptococcus equi subsp. equi, the causative agent of strangles in horses, for the first time. The gene essentiality data for this group C Streptococcus was compared to that of group A and B streptococci.

Results: Six barcoded variants of pGh9:ISS1 were designed and used to generate mutant libraries containing between 33,000-66,000 unique mutants. TraDIS was performed on DNA extracted from each library and data were analysed separately and as a combined master pool. Gene essentiality determined that 19.5% of the S. equi genome was essential. Gene essentialities were compared to those of group A and group B streptococci, identifying concordances of 90.2% and 89.4%, respectively and an overall concordance of 83.7% between the three species.

Conclusions: The use of barcoded pGh9:ISS1 to generate mutant libraries provides a highly useful tool for the assignment of gene function in S. equi and other streptococci. The shared essential gene set of group A, B and C streptococci provides further evidence of the close genetic relationships between these important pathogenic bacteria. Therefore, the ABC of gene essentiality reported here provides a solid foundation towards reporting the functional genome of streptococci.

Item Type: Article
Keywords: Transposon, Sequencing, Essentiality, Barcode
Schools/Departments: University of Nottingham, UK > Faculty of Medicine and Health Sciences > School of Veterinary Medicine and Science
Identification Number: 10.1186/s12864-017-3794-3
Depositing User: Eprints, Support
Date Deposited: 05 Jun 2017 10:31
Last Modified: 05 Jun 2017 10:37
URI: http://eprints.nottingham.ac.uk/id/eprint/43394

Actions (Archive Staff Only)

Edit View Edit View