Staphylococcal infections cause wide spectrum of human illnesses from mild inflammations of skin and soft tissues to life-threatening pneumonia, osteomyelitides and sepses. Treatment of these infections is complicated by emergence both antibiotic-resistant and invasive strains, expressing whole scale of virulence factors. Methicillin-resistant strains, hugely spread in hospital facilities, represent world-wide health risk. Bacteriophages can be utilized as an alternative to antibiotic treatment of bacteria-caused infection, mainly those caused by antibiotic-resistant strains. Description of bacteriophage properties on molecular level is essential for approval of phage-based products in clinical settings. The analysis of factors, which determine ability of bacteriophages to infect Staphylococcus aureus, is precondition for applied research and development of engineered phages directly targeting pathogenic multi-resistant bacteria.

Within presented project I am planning to study structural capsid proteins of polyvalent staphylococcal bacteriophage 812/K1-420, which lyses as much as 90 % of S. aureus strains. The aim of my project is to clone 3 genes encoding different capsid proteins (portal protein, putative tail hydrolase and tail morphogenetic protein) and their subsequent over-expression. The experimental setup is going to involve design of cloning genes’ constructs in pET vectors series, optimizing their over-expression in E. coli culture and isolation of the stable protein forms for their 3D structural analysis using methods of either crystallography or nuclear magnetic resonance. The obtained results will enable further development of modified bacteriophages and legislative approval of studied phage application in the treatment of human infections caused by Staphylococci.

Results

BÁRDY, Pavol, Martin BENEŠÍK, Marta ŠIBOROVÁ, Pavel PLEVKA and Roman PANTŮČEK. Revealing the virion structure of staphylococcal phage 812: Analysis of putative tail hydrolase. Oral presentation, Brno: Masaryk University, 2015. s. 36, 1 s. ISBN 978-80-210-7933-5. BÁRDY, Pavol, Martin BENEŠÍK, Marta ŠIBOROVÁ and Roman PANTŮČEK. Cloning of putative hydrolase gene tmpE of phage 812 and its expression for 3D structural analysis. Poster section, Bratislava: Comenius University in Bratislava, 2015. s. 60-65, 6 s. ISBN 978-80-223-3859-2.

Publications