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Updated: Jun 21, 2026

Using the Overlay Assay to Qualitatively Measure Bacterial Production of and Sensitivity to Pneumococcal Bacteriocins
Published on: September 30, 2014
The streptococcal inhibitor of complement (SIC) protects Streptococcus pyogenes from bacteriocin-like inhibitory
Masaaki Minami1, Daisuke Ohmori, Ichiro Tatsuno
1Department of Bacteriology, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
Abstract:
Streptococcus salivarius inhibits the growth of Streptococcus pyogenes in vitro. Streptococcus pyogenes has various virulence factors, including the streptococcus inhibitor of complement (SIC). Although SIC inhibits the activity of the peptides LL-37 and NAP1, the relationship between SIC and the bacteriocin-like inhibitory substance (BLIS) has not been elucidated. Here, we evaluated whether S. salivarius BLIS affects S. pyogenes SIC. We created three deltasic mutant strains from three S. pyogenes strains and performed deferred antagonism assays. The test strains were BLIS-positive S. salivarius JCM5707 and BLIS-negative S. salivarius NCU12. Deferred antagonism assays with JCM5707 showed that the inhibitory zones in the three deltasic mutant strains were wider than those in the three wild-type strains. Streptococcus pyogenes was cultured in BLIS-containing broth and the change in SIC in the supernatant was assessed by two-dimensional gel electrophoresis (2-DE). The 2-DE analysis of S. pyogenes exoproteins with the JCM5707 supernatant showed reduced SIC compared with those without the JCM5707 supernatant. Changes in sic mRNA levels affected by S. salivarius BLIS were evaluated by a reverse transcriptase-PCR. The sic mRNA level was affected more by the BLIS-positive S. salivarius than by the BLIS-negative strain. Our result indicates that SIC plays a role in the inhibition of S. salivarius BLIS.
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