Targeted and random mutagenesis of Ehrlichia chaffeensis for the identification of genes required for in vivo infection.

ABSTRACT: Ehrlichia chaffeensis is a tick transmitted pathogen responsible for the disease human monocytic ehrlichiosis. Research to elucidate gene function in rickettsial pathogens is limited by the lack of genetic manipulation methods. Mutational analysis was performed, targeting to specific and random insertion sites within the bacterium's genome. Targeted mutagenesis at six genomic locations by homologous recombination and mobile group II intron-based methods led to the consistent identification of mutants in two genes and in one intergenic site; the mutants persisted in culture for 8 days. Three independent experiments using Himar1 transposon mutagenesis of E. chaffeensis resulted in the identification of multiple mutants; these mutants grew continuously in macrophage and tick cell lines. Nine

SUBMITTER: Cheng C 

PROVIDER: S-EPMC3573109 | biostudies-literature | 2013 Feb

REPOSITORIES: biostudies-literature