DEVELOPMENT OF SPECIFIC DNA APTAMERS AGAINST VIRULENCE PROTEASE OF STREPTOCOCCUS PYOGENES
Project no. 661 · Status: Kapanmış · 2016
Summary
Group A Streptococcus is noted for causing necrotizing fasciitis (NF), sepsis and streptococcal toxic shock syndrome, which are known as meat-eating bacterial diseases. SpyCEP is an extracellular protease produced by S. pyogenes. SpyCEP has the ability to disrupt CXC chemokines, and this mechanism of inactivation is related to the invasive character of the infection. In animal models, the spreading rate of SpyCEP deficient S. pyogenes in the host soft tissue has been shown to decrease. DNA aptamers are single-stranded DNA molecules that can bind to their targets with high selectivity and affinity. In similar studies in the literature, DNA aptamers targeting virulent proteins, toxins of various pathogens such as Staphylococcus aureus and Clostridium difficile are available. However, S. pyogenes virulence protein-specific DNA aptamers have been developed for the first time under this project. SELEX (Systematic Evolution of the Ligands by Exponantial Enrichment) technology was used with the aim of developing DNA aptamer that could inhibit SpyCEP enzyme activity. DNA sequencing of the aptamers obtained as a result of SELEX tours was performed and the aptamers were analyzed for preserved motif content. The inhibitory effects of selected aptamers on SpyCEP enzyme activity were investigated. DNA aptamers with inhibitory effect on the enzyme were identified.