Defining the ABC of gene essentiality in streptococci
Autor: | Graham Newland, Duncan J. Maskell, Amelia R. L. Charbonneau, Carl Robinson, Oliver P. Forman, Andrew S. Waller, Amy K. Cain, Mike Boursnell, James A. Leigh, Julian Parkhill |
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Přispěvatelé: | Parkhill, Julian [0000-0002-7069-5958], Maskell, Duncan [0000-0002-5065-653X], Apollo - University of Cambridge Repository, Maskell, Duncan John [0000-0002-5065-653X] |
Jazyk: | angličtina |
Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Transposable element Streptococcus equi lcsh:QH426-470 lcsh:Biotechnology Mutant Genomics Biology Genome DNA sequencing 03 medical and health sciences Transposon Sequencing Essentiality Barcode lcsh:TP248.13-248.65 Genetics Sequencing Gene Transposon Barcode Genes Essential High-Throughput Nucleotide Sequencing Streptococcus Essentiality lcsh:Genetics 030104 developmental biology Essential gene Genes Bacterial Mutation Biotechnology Research Article |
Zdroj: | BMC Genomics, Vol 18, Iss 1, Pp 1-11 (2017) BMC Genomics |
ISSN: | 1471-2164 |
Popis: | 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. Electronic supplementary material The online version of this article (doi:10.1186/s12864-017-3794-3) contains supplementary material, which is available to authorized users. |
Databáze: | OpenAIRE |
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