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Genetic transformation of Gardnerella species and characterization of vaginolysin and sialidase mutants | Infection and Immunity
Genetic manipulation is a fundamental process in the study of bacterial pathogenesis. The ability to make mutations and complements is necessary for definitive identification of virulence genes, and the ability to make unmarked mutations is preferable in these studies (1). Certain bacterial pathogens were considered genetically intractable for many years, such as Mycobacterium tuberculosis or Chlamydia trachomatis, and studies of their virulence mechanisms necessarily focused on epidemiology, immunology, cell biology in the host, and proteomic or transcriptomic analyses (2, 3). Those classic cases were eventually resolved by the discovery of effective transduction, natural transformation, and chemical transformation methods, greatly facilitating the identification of virulence factors (4–7). Gardnerella species have similarly been considered genetically intractable, and like M. tuberculosis and C. trachomatis, Gardnerella have an unusual cell envelope. Similar to other Actinobacteria, Gardnerella are generally considered gram-positive, but stain gram-variable because of their unusual and incompletely characterized envelope (8).
