Development of an in vivo Himar1 transposon mutagenesis system for use in Streptococcus equi subsp. equi.
Streptococcus equi subsp. equi is the causative agent of the equine disease strangles. In this study we describe the development of an in vivo Himar1 transposon system for the random mutagenesis of S. equi and, potentially, other Gram-positive bacteria. We demonstrate efficient and random transposition of a modified mini-transposon onto the chromosome by Southern blot analysis and insertion site sequencing. Non-haemolytic mutants were isolated at a frequency of 0.2%, and acapsular mutants at a frequency of 0.04%. Taken together, these data demonstrate that in vivo Himar1 mutagenesis can be used for genomic-scale mutational analysis of S. equi, and is likely to be applicable to the study of other streptococci.
May
James P
JP
Department of Clinical Veterinary Medicine, Centre for Veterinary Science, University of Cambridge, Madingley Road, Cambridge CB3 0ES, UK.
Walker
Caray A
CA
Maskell
Duncan J
DJ
Slater
Josh D
JD
D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D018274:Electroporation; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics; D016254:Mutagenesis, Insertional / Q000379:methods; D010957:Plasmids; D018502:Streptococcus equi / Q000737:chemistry / Q000235:genetics*; D017011:Streptococcus suis / Q000737:chemistry / Q000235:genetics*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial