ABSTRACT
A 0.8-kb HindIII-BamHI internal fragment of the Streptococcus mutans wall-associated protein A gene (wapA) was ligated to the 5.1-kb HindIII-BamHI fragment of the chimeric Streptococcus-Escherichia coli plasmid pVA891 (Emr Cmr). The resulting construct was used to transform S. mutans GS-5, and erythromycin-resistant mutants were isolated and analyzed. Directed mutagenesis of the wapA gene by plasmid insertion through homologous recombination was demonstrated by Southern blot hybridization with the wapA and pVA891 probes. Stable mutants were obtained, and the alteration of the wapA gene by insertional inactivation was associated with a significant decrease in S. mutans sucrose-dependent aggregation and binding to smooth surfaces. Thus, WapA may play an important role in the colonization of the tooth surface by S. mutans and in the buildup of dental plaque. These findings provided an explanation for previous studies which indicated that WapA was effective in the prevention of dental caries in animal models. Thus, the use of recombinant WapA in the preparation of a safe and effective human dental vaccine should be investigated further.
