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Generation of a CRISPR database for Y ersinia pseudotuberculosis complex and role of CRISPR ‐based immunity in conjugation
Author(s) -
Koskela Katja A.,
Mattinen Laura,
KalinMänttäri Laura,
Vergnaud Gilles,
Gorgé Olivier,
Nikkari Simo,
Skurnik Mikael
Publication year - 2015
Publication title -
environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.954
H-Index - 188
eISSN - 1462-2920
pISSN - 1462-2912
DOI - 10.1111/1462-2920.12816
Subject(s) - crispr , biology , plasmid , genetics , gene , direct repeat , dna , palindrome , corynebacterium pseudotuberculosis , trans activating crrna , microbiology and biotechnology , bacteria , genome
Summary The clustered regularly interspaced short palindromic repeat – CRISPR‐associated genes ( CRISPR‐C as) system is used by bacteria and archaea against invading conjugative plasmids or bacteriophages. Central to this immunity system are genomic CRISPR loci that contain fragments of invading DNA . These are maintained as spacers in the CRISPR loci between direct repeats and the spacer composition in any bacterium reflects its evolutionary history. We analysed the CRISPR locus sequences of 335 Y ersinia pseudotuberculosis complex strains. Altogether 1902 different spacer sequences were identified and these were used to generate a database for the spacer sequences. Only ∼10% of the spacer sequences found matching sequences. In addition, surprisingly few spacers were shared by Y ersinia pestis and Y . pseudotuberculosis strains. Interestingly, 32 different protospacers were present in the conjugative plasmid pYptb 32953. The corresponding spacers were identified from 35 different Y . pseudotuberculosis strains indicating that these strains had encountered pYptb 32953 earlier. In conjugation experiments, pYptb 32953‐specific spacers generally prevented conjugation with spacer‐positive and spacer‐free strains. However, some strains with one to four spacers were invaded by pYptb 32953 and some spacer‐free strains were fully resistant. Also some spacer‐positive strains were intermediate resistant to conjugation. This suggests that one or more other defence systems are determining conjugation efficiency independent of the CRISPR‐Cas system.