Genetic transformation of species and characterization of vaginolysin and sialidase mutants.

Bacterial vaginosis (BV) is the most prevalent vaginal disorder in women of childbearing age and causes pregnancy complications including preterm birth. Species of increase just prior to the onset of symptoms and are considered to play major roles in the development and transmission of BV. However, species have remained genetically intractable, limiting investigations of their virulence mechanisms. Here we describe methods for genetic manipulation of . Through trial and error we optimized methods for electrotransformation of and created methods for making mutations and complements. We mutated the gene for the toxin vaginolysin () in and the gene for sialidase in . A point mutant was tested in human cervix tissue and found to lack lytic activity. The mutant lost sialidase and mucus degradation activity. Overall, this genetic toolkit opens a door for molecular characterization of and its mechanisms in BV.