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Microbiology 151 (2005), 3947-3955; DOI  10.1099/mic.0.28221-0
© 2005 Society for General Microbiology

Identification of genes associated with mutacin I production in Streptococcus mutans using random insertional mutagenesis

Phoebe Tsang1, Justin Merritt2, Trang Nguyen1, Wenyuan Shi1,2 and Fengxia Qi1

1 UCLA School of Dentistry, Los Angeles, CA 90025, USA
2 UCLA Molecular Biology Institute, Los Angeles, CA 90025, USA

Correspondence
Fengxia Qi
fqi{at}dent.ucla.edu

Streptococcus mutans is a major pathogen implicated in dental caries. Its virulence is enhanced by its ability to produce bacteriocins, called mutacins, which inhibit the growth of other Gram-positive bacteria. The goal of this study is to use a random insertional mutagenesis approach to search for genes that are associated with mutacin I production in the virulent strain UA140. A random insertional mutagenesis library consisting of 11 000 clones was constructed and screened for a mutacin-defective phenotype. Mutacin-defective clones were isolated, and their insertion sites were determined by PCR amplification or plasmid rescue followed by sequencing. A total of twenty-five unique genes were identified. These genes can be categorized into the following functional classes: two-component sensory systems, stress responses, energy metabolism and central cellular processes. Several conserved hypothetical proteins with unknown functions were also identified. These results suggest that mutacin I production is stringently controlled by diverse and complex regulatory pathways.

Abbreviations: PTS, phosphoenolpyruvate : sugar phosphotransferase system; TCS, two-component sensory system




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